SEARCH

SEARCH BY CITATION

Keywords:

  • Alzheimer disease;
  • amyotrophic lateral sclerosis;
  • cytoskeleton;
  • energy metabolism;
  • Huntington disease;
  • mitochondria;
  • neurodegeneration;
  • nitration;
  • oxidative stress;
  • Parkinson disease;
  • tauopathies

Abstract

  1. Top of page
  2. Abstract
  3. INTRODUCTION
  4. IDENTIFICATION OF OXIDIZED AND NITRATED PROTEINS
  5. ALZHEIMER DISEASE
  6. TAUOPATHIES
  7. PARKINSON DISEASE AND RELATED α-SYNUCLEINOPATHIES
  8. HUNTINGTON DISEASE
  9. AMYOTROPHIC LATERAL SCLEROSIS
  10. COMMON AND SPECIFIC PROTEIN TARGETS IN DIFFERENT PATHOLOGIES
  11. OXIDATIVE CHANGES IN POST-MORTEM BRAIN ARE PRINCIPALLY PRIMARY
  12. PROTEIN OXIDATION IS AN EARLY EVENT IN NEURODEGENERATIVE DISEASES
  13. OXIDATIVE DAMAGE AND LOSS OF FUNCTION
  14. CLINICAL IMPLICATIONS
  15. CELLULAR LOCALIZATION OF OXIDATIVELY DAMAGED PROTEINS
  16. PITFALLS AND LIMITATIONS
  17. REFINING METHODS TO IMPROVE REDOX PROTEOMICS
  18. CONCLUDING COMMENTS
  19. ACKNOWLEDGMENTS
  20. REFERENCES
  21. Supporting Information

Human neurodegenerative diseases with abnormal protein aggregates are associated with aberrant post-translational modifications, solubility, aggregation and fibril formation of selected proteins which cannot be degraded by cytosolic proteases, ubiquitin–protesome system and autophagy, and, therefore, accumulate in cells and extracellular compartments as residual debris. In addition to the accumulation of “primary” proteins, several other mechanisms are involved in the degenerative process and probably may explain crucial aspects such as the timing, selective cellular vulnerability and progression of the disease in particular individuals. One of these mechanisms is oxidative stress, which occurs in the vast majority of, if not all, degenerative diseases of the nervous system. The present review covers most of the protein targets that have been recognized as modified proteins mainly using bidimensional gel electrophoresis, Western blotting with oxidative and nitrosative markers, and identified by mass spectrometry in Alzheimer disease; certain tauopathies such as progressive supranuclear palsy, Pick disease, argyrophilic grain disease and frontotemporal lobar degeneration linked to mutations in tau protein, for example, FTLD-tau, Parkinson disease and related α-synucleinopathies; Huntington disease; and amyotrophic lateral sclerosis, together with related animal and cellular models. Vulnerable proteins can be mostly grouped in defined metabolic pathways covering glycolysis and energy metabolism, cytoskeletal, chaperoning, cellular stress responses, and members of the ubiquitin–proteasome system. Available information points to the fact that vital metabolic pathways are hampered by protein oxidative damage in several human degenerative diseases and that oxidative damage occurs at very early stages of the disease. Yet parallel functional studies are limited and further work is needed to document whether protein oxidation results in loss of activity and impaired performance. A better understanding of proteins susceptible to oxidation and nitration may serve to define damaged metabolic networks at early stages of disease and to advance therapeutic interventions to attenuate disease progression.


INTRODUCTION

  1. Top of page
  2. Abstract
  3. INTRODUCTION
  4. IDENTIFICATION OF OXIDIZED AND NITRATED PROTEINS
  5. ALZHEIMER DISEASE
  6. TAUOPATHIES
  7. PARKINSON DISEASE AND RELATED α-SYNUCLEINOPATHIES
  8. HUNTINGTON DISEASE
  9. AMYOTROPHIC LATERAL SCLEROSIS
  10. COMMON AND SPECIFIC PROTEIN TARGETS IN DIFFERENT PATHOLOGIES
  11. OXIDATIVE CHANGES IN POST-MORTEM BRAIN ARE PRINCIPALLY PRIMARY
  12. PROTEIN OXIDATION IS AN EARLY EVENT IN NEURODEGENERATIVE DISEASES
  13. OXIDATIVE DAMAGE AND LOSS OF FUNCTION
  14. CLINICAL IMPLICATIONS
  15. CELLULAR LOCALIZATION OF OXIDATIVELY DAMAGED PROTEINS
  16. PITFALLS AND LIMITATIONS
  17. REFINING METHODS TO IMPROVE REDOX PROTEOMICS
  18. CONCLUDING COMMENTS
  19. ACKNOWLEDGMENTS
  20. REFERENCES
  21. Supporting Information

In most aerobic cell types the mitochondrial respiratory chain is one of the main sources of generation of reactive oxygen species (ROS) under physiologic conditions (6, 45, 46, 68, 83, 112). In addition to mitochondria, peroxisomes, endoplasmatic reticulum, microsomes, nucleus and plasma membrane oxidases are potential sources of ROS. Univalent oxygen reduction by the mitochondrial respiratory chain, as well as metal-ion-catalyzed reactions, generates a wide diversity of highly reactive metabolites of oxygen and nitrogen. These products mainly include superoxide anion (O-2), hydrogen peroxide (H2O2), hydroxyl radical (HO·), which can be formed from either the O-2 and H2O2 (Haber–Weiss reaction) or from metal ion (Fe2+, Fe3+) and H2O2 (Fenton reaction), peroxyl radical (RO·2), alkoxyl radical (RO·), hydroperoxyl radical (HO·2), hypochlorous acid (HOCl), hypobromous acid (HOBr) and singlet oxygen (O2) (50).

ROS plays a vital signaling role in physiologic conditions (50, 66, 120). However, ROS surpassing antioxidant cellular stress responses can be considered a significant source of endogenous structural damage to other cellular macromolecules, including DNA, RNA, carbohydrates, lipids and proteins, finally producing cytotoxic effects.

Nitric oxide (NO·) is produced by the oxidation of one of the terminal guanidonitrogen atoms of l-arginine catalyzed by different isoforms of nitric oxide synthase. NO plays a crucial role in physiologic conditions, such as autoimmunity, muscular relaxation and neurotransmission.

Nevertheless, NO· is also a source of harmful reactive nitrogen species (RNS). Main RNS are nitrogen dioxide (·NO2), nitrous acid (HNO2), nitrosyl cation (NO+), nitrosyl anion (NO-), dinitrogen tetroxide (N2O4), dinitrogen trioxide (N2O3), peroxynitrite (ONOO-), peroxynitrous acid (ONOOH), alkyl peroxynitrites (ROONO), nitronium cation (NO+2) and nitryl chloride (NO2Cl) (66).

All amino acid residues are susceptible to oxidation, but ion-catalyzed oxidation of some residues may result in the production of protein carbonyl derivatives (37, 108, 111). Characteristic products are glutamic semialdehyde and aminoadipic semialdehyde, which are derived from arginine/proline and lysine, respectively (38, 102). Because the magnitude of protein carbonylation is higher than any other primary change resulting from oxidation, carbonylation of proteins is currently used as a marker of protein oxidation in variegated settings (1, 15, 35, 60, 110).

In addition to direct effects, protein oxidative modifications may also occur following the reaction of distinct reactive carbonyl species (RCS) as glyoxal, glycoaldehyde, methylglyoxal, malondialdehyde (MDA) and 4-hydroxynonenal (HNE), derived from the oxidation of carbohydrates and lipids. Carbonyl species react with lysine, arginine and cysteine residues leading to the formation of advanced glycation and lipoxidation end-products (AGE/ALEs) in proteins. Typical AGEs/ALEs adducts are MDA-lysine (MDAL), carboxymethyl-lysine (CML) and carboxyethyl-lysine (CEL), among many others (82, 85, 122, 124).

Regarding RNS, NO damage to thiols, amines and hydroxyls leads to nitrosative damage. Reactions with RNS lead to the formation of 3-nitrotyrosine (nitration) and to oxidation of distinct substrates. As an example, reactive peroxinitrite is able to nitrate tyrosine residues and to oxidize methionine residues of proteins (55, 99, 121).

Cells have developed different mechanisms to prevent oxidative molecular damage. Antioxidant enzymes are superoxide dismutases, including cytosolic Cu,Zn-superoxide dismutase (SOD1), matrix mitochondrial Mn-superoxide dismutase (SOD2) and extracellular superoxide dismutase 3 (SOD3); catalase; glutathione peroxidase; peroxiredoxin; and some molecular chaperones. Non-enzymatic systems composed of different proteins such as ferritin (binds iron in the cytoplasm of mammalian cells) and ceruloplasmin (binds copper in plasma) have the capacity to bind transition metals in oxidation reactions. Finally, α-tocopherol (vitamin E), ascorbic acid (vitamin C), glutathione (l-γ-glutamyl-l-cysteinyl-glycine), flavonoids and carotenoids may act as antioxidants (50).

The concept of oxidative stress has been applied to the imbalance between the generation of ROS/RNS/RCS, and the cellular antioxidant defense mechanisms (4, 49). This may result in oxidative damage to varied molecules including DNA, RNA, lipids and proteins. Oxidative damage increases in aging (15, 47, 60, 97, 109–111). The nervous system is particularly susceptible to oxidative stress because of the abundance of Polyunsaturated fatty acids (PUFA) content, especially arachidonic and docosahexaenoic acids, the high oxygen consumption rate, and the relatively low levels of antioxidant pathways (7, 14, 31). The presence of increased oxidative stress and oxidative damage in neurodegenerative diseases has been recognized for years, and it has been the subject of hundreds of papers and reviews (25, 39, 41, 44, 61–63, 67, 75, 80, 81, 90, 98, 113, 127–129). However, little is known about the specific protein targets of oxidative damage in human neurodegenerative diseases.

The term proteomics is used to define the analysis of the whole proteins expressed by a genome. Redox proteomics is used to name the analysis of proteins modified by oxidation and nitration (16, 34). The term redox proteomics is instrumental as it serves to identify proteins that are damaged as a result of oxidation as well as the methods used to recognize modified proteins (36).

The objectives of the present review are (i) to list proteins modified by oxidation/nitration identified so far in neurodegenerative diseases covering Alzheimer disease (AD), tauopathies, Parkinson disease (PD) and related α-synucleinopathies, Huntington disease (HD), and amyotrophic lateral sclerosis (ALS), and related animal and cellular models; (ii) to give information about the methods used to identify those proteins in the different studies; (iii) to identify vulnerable metabolic pathways in individual diseases and vulnerable proteins common to different neurodegenerative disorders; (iv) to investigate the effects of oxidative stress on protein targets at early stages of neurodegenerative diseases to learn whether oxidative damage to proteins is an early event in degenerative diseases of the nervous system; (v) to find out whether studies dealing with protein damage resulting from oxidation/nitration have been accompanied by studies focused on associated loss of function; (vi) to clarify whether information is available regarding the involvement of particular cell types; (vii) to discuss limitations of redox proteomics; and (viii) to comment on aspects that may help to improve the use and results of redox proteomics applied to the study of neurodegenerative diseases.

IDENTIFICATION OF OXIDIZED AND NITRATED PROTEINS

  1. Top of page
  2. Abstract
  3. INTRODUCTION
  4. IDENTIFICATION OF OXIDIZED AND NITRATED PROTEINS
  5. ALZHEIMER DISEASE
  6. TAUOPATHIES
  7. PARKINSON DISEASE AND RELATED α-SYNUCLEINOPATHIES
  8. HUNTINGTON DISEASE
  9. AMYOTROPHIC LATERAL SCLEROSIS
  10. COMMON AND SPECIFIC PROTEIN TARGETS IN DIFFERENT PATHOLOGIES
  11. OXIDATIVE CHANGES IN POST-MORTEM BRAIN ARE PRINCIPALLY PRIMARY
  12. PROTEIN OXIDATION IS AN EARLY EVENT IN NEURODEGENERATIVE DISEASES
  13. OXIDATIVE DAMAGE AND LOSS OF FUNCTION
  14. CLINICAL IMPLICATIONS
  15. CELLULAR LOCALIZATION OF OXIDATIVELY DAMAGED PROTEINS
  16. PITFALLS AND LIMITATIONS
  17. REFINING METHODS TO IMPROVE REDOX PROTEOMICS
  18. CONCLUDING COMMENTS
  19. ACKNOWLEDGMENTS
  20. REFERENCES
  21. Supporting Information

Several methods are currently used to identify oxidative stress and oxidative damage in tissues (36, 37). Yet the majority of studies dealing with the identification of proteins modified by oxidation in human neurodegenerative diseases are based on bidimensional gel electrophoresis of paired gels run in parallel—one of them is used to transfer proteins to membranes to carry out Western blotting with specific oxidative damage markers, and the other serves to pick up selected spots for protein identification by mass spectrometry. Common antibodies utilized to identify modified proteins are anti-AGE, anti-CEL, anti-CML, anti-3-NTyr, anti-MDAL and anti-HNE. The recognition of spots of modified proteins in Western blots is conducted in the parallel gels stained with Coomassie blue, SYPRO Ruby or silver. This is followed by in-gel digestion of the selected spots, analysis of fingerprints by MALDI mass spectrometry and identification of proteins using a database. To detect carbonyls, samples are derivatized to hydrazones with 2,4-dinitrophenylhydrazyne (DNPH) usually before the separation of proteins.

Several variables are introduced in different studies, including the characteristics of buffers of homogenates and the solutions used for protein loading. These aspects may have implications on the range and type of proteins finally transferred to membranes for Western blotting labeling (see section Pitfalls and limitations).

The effects of post-mortem delay in the study of oxidized/nitrated proteins in human brain has been analyzed by freezing part of the sample immediately 1 or 2 h after death, and storing pieces of the remaining sample at −4°C (thus mimicking corpse preservation) and then freezing them at 2, 6, 8, 12, 18, 24 and 48 h at –80°C until use. Monodimensional gel electrophoresis and Western blotting to anti-MDAL, anti-HNE, anti-CEL, anti-CML and anti-3-NTyr antibodies has demonstrated good preservation up to 12–18 h. Yet reduction or enhancement of the intensity of previous bands and appearance of new bands occurs from this time onward (43).

The relevant methodological aspects in individual studies, including use of total homogenates or subfractions, buffers, regions examined, gel staining, methods employed for mass spectrometry and software characteristics, are shown in Tables S1–S3 (Supporting Information).

ALZHEIMER DISEASE

  1. Top of page
  2. Abstract
  3. INTRODUCTION
  4. IDENTIFICATION OF OXIDIZED AND NITRATED PROTEINS
  5. ALZHEIMER DISEASE
  6. TAUOPATHIES
  7. PARKINSON DISEASE AND RELATED α-SYNUCLEINOPATHIES
  8. HUNTINGTON DISEASE
  9. AMYOTROPHIC LATERAL SCLEROSIS
  10. COMMON AND SPECIFIC PROTEIN TARGETS IN DIFFERENT PATHOLOGIES
  11. OXIDATIVE CHANGES IN POST-MORTEM BRAIN ARE PRINCIPALLY PRIMARY
  12. PROTEIN OXIDATION IS AN EARLY EVENT IN NEURODEGENERATIVE DISEASES
  13. OXIDATIVE DAMAGE AND LOSS OF FUNCTION
  14. CLINICAL IMPLICATIONS
  15. CELLULAR LOCALIZATION OF OXIDATIVELY DAMAGED PROTEINS
  16. PITFALLS AND LIMITATIONS
  17. REFINING METHODS TO IMPROVE REDOX PROTEOMICS
  18. CONCLUDING COMMENTS
  19. ACKNOWLEDGMENTS
  20. REFERENCES
  21. Supporting Information

Excellent reviews of oxidatively damaged proteins in AD and related models, identified by researchers of the University of Kentucky, have recently appeared (20, 118). Yet other groups have significantly contributed to identify proteins modified by oxidation. For these reasons, the present review updates and complements the list by adding important observations made in other centers. To facilitate understanding, vulnerable proteins have been grouped in defined metabolic pathways covering glycolysis and energy metabolism, mitochondrial electron transport chain and oxidative phosphorylation, structural proteins, chaperones, stress proteins, ubiquitin–proteasome system components, and other proteins.

Glycolysis and energy metabolism

Aldolase A, which catalyzes d-fructose 1,6-bisphosphate to glyceraldehyde 3-phosphate and dihydroxyacetone phosphate, is modified by oxidation and nitration at middle and advanced stages of AD (57, 101); oxidative damage to aldolase C has also been detected in advanced stages of AD (58). Triose phosphate isomerase, which catalyzes the reversible interconversion of glyceraldehyde 3-phosphate and dihydroxyacetone phosphate, is also modified by oxidation and nitration at middle and advanced stages (24, 101, 115). Glyceraldehyde 3-phosphate dehydrogenase (GAPDH), involved in the two-step reaction that transforms glyceradehyde 3-phosphate to d-glycerate 1,3-bisphosphate, is modified by S-glutathionylation (77) and nitration (117) in AD.

Interestingly, GAPDH is carbonylated in the brain of Wistar rats following intracerebral injection of amyloid-beta1–42 (Aβ1–42) (12), and in neuronal cultures treated with Aβ1–42 (116).

Phosphoglycerate kinase (PGK) catalyzes the reaction of d-glycerate 1,3-bisphosphate with adenosine diphosphate (ADP) to form adenosine triphosphate (ATP) and 3-phosphoglycerate. Oxidized PGK, as detected using anti-4-HNE, has been shown at middle stages of AD-related pathology to be clinically manifested as mild cognitive impairment (100).

Phosphoglycerate mutase (PGM) catalyzes an internal transfer of a phosphate group from 3-phosphoglycerate to 2-phosphoglycerate. Isoform B (PGM-1) is more oxidized and nitrated in AD cases than in age-matched controls (101, 115).

Similarly, PGM-1 was more carbonylated in 3-month-old male Wistar rats following intracerebral injection of Aβ1–42 when compared with controls (12).

Enolases, which modulate the reaction of 2-phosphoglycerate to phosphoenolpyruvate in the next-to-last step of glycolysis, are targets of oxidative and nitrosative damage in AD. α-Enolase has increased carbonylation, lipoxidation, S-glutathionylation and nitration levels in advanced stages of AD and in cases of mild cognitive impairment (19, 23, 24, 77, 84, 100, 101, 114, 115). Likewise, increased oxidized α-enolase has been found in the brain of mutant Tg2576 mice which bear the Swedish APP mutation causative of familial AD (105).

Increased modification of enolase, as revealed with anti-DNPH, anti-MDAL and anti-3NTyr antibodies, has been described in sporadic AD and in cases with familial AD linked with mutations in presenilin-1 (18, 24, 84).

The last enzyme of glycolysis is pyruvate kinase (PK), which catalyzes the step from phosphoenolpyruvate to pyruvate, thus transferring phosphate to ADP to form ATP. Increased oxidation of isoform PK-M2 was reported in human brain samples of cases with mild cognitive impairment by using anti-DNPH and anti-HNE antibodies (19, 100). In the same line, increased PK oxidation occurred in neuronal cultures from rat fetuses exposed to Aβ1–42 (116).

Two enzymes of the Krebs cycle appear to be targets of oxidation in AD and related models. Pyruvate dehydrogenase, which catalyzes the step from pyruvate to acetyl-CoA, is more carbonylated in the brain of rats treated with intracerebral injection of Aβ1–42 (12). Malate dehydrogenase, which catalyzes the interconversion of malate and oxaloacetate using nicotinamide adenine as a coenzyme, has increased levels of carbonylation in primary rat neuronal cultures treated with Aβ1–42 (116). Moreover, similar results have been obtained in transgenic Caenorhabditis elegans (13).

In addition to enzymes involved in glycolysis and Krebs cycle, several proteins linked to variegated metabolic reactions have been shown to be targets of oxidative damage in AD. These include carbonyl reductase 1, an oxoreductase enzyme related to arachidonic acid metabolism (100), carbonyl anhydrase (58), carbonic anhydrase II (CA II) (114, 117), and glutamate dehydrogenase, which converts glutamate to α-ketoglutarate (101). Increased carbonyl levels in GDH have also been discovered in gerbil synaptosomes exposed to Aβ1–42 (11).

Related to nitrogen metabolism, glutamine synthetase (GS), which catalyzes glutamate and ammonia to form glutamine, is more oxidized in AD cases when compared with age-matched controls (19, 22). Increased oxidation of glutamate-ammonia ligase, a transferase enzyme, occurs after intracerebral injection of Aβ1–42 in rat brain (12). Finally, lactate dehydrogenase B (LDH 2), which participates in the interconversion of pyruvate and lactate, is more oxidized in cases of AD presenting as mild cognitive impairment compared with controls (100).

Electron transport chain, oxidative phosphorylation and other mitochondrial components

Complex V or ATP synthase catalyzes the synthesis of ATP from ADP and inorganic phosphate with a flow of protons from the intermembrane space to the matrix side. Several studies have found lipoxidized and nitrated ATP synthase in middle and advanced stages of AD (84, 100, 117). ATP synthase oxidative damage is a very early event in AD, as ATP synthase has been found oxidized and its function reduced in the entorhinal cortex in asymptomatic cases with Braak II AD-related pathology (119).

In the same line, ATP synthase is more carbonylated in stable transgenic C. elegans strain CL 2337 when compared with wild worms (13).

Ubiquinol–cytochrome c reductase complex core protein I is a component of the complex III, which helps to link the complex between cytochromes c and c1. This protein is more lipoxidated in the frontal cerebral cortex of advanced AD when compared with control samples (84).

Evidence of increased oxidative damage in creatine kinase BB (CK BB) derives from the observation that specific protein carbonyl content is higher in AD cases when compared with controls (2, 3, 22), and in AD-related mice models (26).

Voltage-dependent anion-channel protein-1 (VDAC-1) is a porin that forms a channel through the mitochondrial outer membrane and the plasma membrane. It helps the transport of a variety of purine nucleotides (responsible for ATP/ADP exchange) and allows the diffusion of small hydrophilic molecules. VDAC-1 also has an important role as a regulator of mitochondrial function. Nitrated VDAC-1 is significantly increased in AD (117).

Structural proteins

Cytoskeletal proteins are targets of oxidative damage in AD. Increased β-actin carbonylation has been found in sporadic AD and in familial AD due to mutations in presenilin-1 (3, 18). Similar changes occur at earlier stages of AD corresponding with clinical symptoms of mild cognitive impairment (100). Transgenic C. elegans expressing human Aβ1–42 also show β-actin oxidative damage (13). Increased β-actin oxidation also occurs in synaptosomes of Mongolian gerbils exposed to Aβ1–42 (11, 12).

Oxidative damage to α-tubulin 1, as revealed with anti-MDAL antibodies, has been reported in AD (84). Tubulins are also targets of oxidative damage in the brains of rats following intracerebral injection of Aβ1–42 (12).

By using a different approach, it has been shown that high molecular neurofilament proteins are substrates of adduction by HNE (123). Carbonyl-related modifications of neurofilament protein have been shown in neurofibrillary tangles in AD (106).

Glial fibrillary acidic protein (GFAP) is oxidized in the normal aged brain, but GFAP oxidative damage increases in AD (57, 84) and related animal models (11).

Chaperones, stress proteins and stress responses

There is cumulative evidence of increased oxidation of several chaperones including HSC-71 (23) and HSP-70 (100) in AD, and HSP-60 in experimental models (12, 26). αB-crystallin is also a target of S-glutathionylation in AD (77).

Pin-1 is a protein within the peptidyl-prolyl isomerase family with chaperone activity involved in several cellular functions, including the modulation of assembly and folding of several proteins. Increased oxidized Pin-1 level, using anti-DNPH antibodies, has been found in the hippocampus in AD (114).

Regarding oxidative stress responses, SOD1 is oxidatively damaged in AD (28).

Ubiquitin–proteasome system

Ubiquitin carboxy-terminal hydrolase L-1 (UCHL-1) belongs to a family of proteases with high specificity for ubiquitinated substrates. Increased levels of carbonylated and oxidized UCHL-1 have been reported in AD (22). Oxidative modifications of UCHL-1 in AD have also been reported by independent groups (27).

Additional targets of oxidative damage in AD

Other proteins are oxidatively damaged or are targets of nitration in AD cases. Most of the descriptions refer to unique reports that should be validated by further studies. Moreover, some of them appear as isolated molecules within a particular metabolic pathway. A list of altered proteins in AD is shown in Table 1. In addition to AD, details are also provided for several experimental models including intracerebral injection of Aβ1–42 in rats and gerbils, transgenic mice, transgenic C. elegans and transfected cell lines.

Table 1.  Alzheimer disease.
ReferenceDisease/stageProteinFunctionDetection antibodyDegree of oxidation in relation to age-matched controlsEnzymatic activityTotal protein levelsComments
Aksenov et al (2)ADCK BBEnergy transductionAnti-DNPH antibody[UPWARDS ARROW][DOWNWARDS ARROW][DOWNWARDS ARROW]
Aksenov et al (3)ADβ-tubulinCytoskeletonAnti-DNPH antibody=NANA
 β-actinCytoskeleton[UPWARDS ARROW]NANA
CK BBEnergy transduction[UPWARDS ARROW]NANA
Castegna et al (22)ADCK BBEnergy transductionAnti-DNPH antibody[UPWARDS ARROW]NANA
 UCHL-1UPS[UPWARDS ARROW]NANA
GSAmino acid biosynthesis[UPWARDS ARROW]NANA
Castegna et al (23)ADDRP-2Pyrimidine metabolismAnti-DNPH antibody[UPWARDS ARROW]NANA
 α-enolaseGlycolysis[UPWARDS ARROW]NANA
HSC-71Chaperone[UPWARDS ARROW]NANA
Castegna et al (24)ADα-EnolaseGlycolysisAnti-3-NTyr antibody[UPWARDS ARROW]NANA
 Triosephosphate isomeraseGlycolysis[UPWARDS ARROW]NANA
Neuropolypeptide h3Phospholipid binding protein[UPWARDS ARROW]NANA
β-actinCytoskeleton=NANA
LDHGlycolysis=NANA
γ-enolaseGlycolysis=NANA
Choi et al (27)ADUCH-L1UPSAnti-DNPH antibody[UPWARDS ARROW]NA[DOWNWARDS ARROW] 50%identification of oxidation sites
Choi et al (28)ADSOD1Antioxidant responseAnti-DNPH antibody[UPWARDS ARROW]NA[UPWARDS ARROW]IHC
Pamplona et al (84)AD Stages V-VI/CNeurofilament triplet LCytoskeletonAnti-MDAL-antibody[UPWARDS ARROW] (3-fold)NANA
 VimentinCytoskeleton=NANA
β-tubulin 2Cytoskeleton=NANA
α-tubulin 1Cytoskeleton[UPWARDS ARROW] (9-fold)NANA
α-tubulin 4Cytoskeleton=NANA
α-tubulin 6Cytoskeleton=NANA
β-actinCytoskeleton=NANA
γ-actinCytoskeleton=NANA
GFAPCytoskeleton[UPWARDS ARROW] (8-fold)NANA
γ-enolaseGlycolysis[UPWARDS ARROW] (2-fold)NANA
α-enolaseGlycolysis=NANA
Ubiquinol-cytochrome c reductase complex core protein IElectron transport chain[UPWARDS ARROW] (4-fold)NANA
ATP synthase (β chain)Oxidative phosphorylation[UPWARDS ARROW] (4-fold)NANA
CK BBEnergy transduction=NANA
GSAmino acid biosynthesis=NANA
Glutamate dehydrogenase 1Amino acid biosynthesis=NANA
Guanine nucleotide-binding protein G(I)/G(S)/G(T) βSignal transduction=NANA
60-kDa HSPChaperone=NANA
Dihydropyrimidinase-related protein-2Pyrimidine metabolism=NANA
Korolainen et al (57)ADGFAPCytoskeletonAnti-DNPH antibody[UPWARDS ARROW]NA[UPWARDS ARROW]
 α-actinCytoskeleton [UPWARDS ARROW]NANA
β-actinCytoskeleton[UPWARDS ARROW]NANA
EnolaseGlycolysis[UPWARDS ARROW]NANA
CK BBEnergy transduction[UPWARDS ARROW]NANA
Korolainen et al (58)AD Braak stages V-VICA IICO2 metabolismAnti-DNPH antibody[UPWARDS ARROW]NA=Oxp/tp =
 MDH1aKreb's cycle[DOWNWARDS ARROW]NA[UPWARDS ARROW]Oxp/tp [DOWNWARDS ARROW]
MDH1bKreb's cycle[DOWNWARDS ARROW]NA=Oxp/tp [DOWNWARDS ARROW]
AconitaseKreb's cycle[DOWNWARDS ARROW]NA=Oxp/tp =
Glutamate dehydrogenaseAmino acid biosynthesis=NA=Oxp/tp [DOWNWARDS ARROW]
14-3-3 protein zeta/deltaCell signaling[DOWNWARDS ARROW]NA=Oxp/tp =
Aldolase CGlycolysis[DOWNWARDS ARROW]NA=Oxp/tp =
Aldolase AGlycolysis[DOWNWARDS ARROW]NA=Oxp/tp =
ATP synthaseOxidative phosphorylation=NA[UPWARDS ARROW]Oxp/tp =
Butterfield et al (19)AD MCI Braak stages III, V, VIEnolase 1GlycolysisAnti-DNPH antibody[UPWARDS ARROW][DOWNWARDS ARROW]NAIP, IHC
 GSEnergy transduction[UPWARDS ARROW][DOWNWARDS ARROW]NAIP, IHC
Pyruvate kinase M2Glycolysis[UPWARDS ARROW][DOWNWARDS ARROW]NA
PIN 1Chaperone[UPWARDS ARROW][DOWNWARDS ARROW]NA
Butterfield et al (18)Familial AD with mutations in PS1γ-enolaseGlycolysisAnti-DNPH antibody[UPWARDS ARROW]NANA
 ActinCytoskeleton[UPWARDS ARROW]NANA
DMDMAH-1Nitric oxide metabolism[UPWARDS ARROW]NANA
UCHL-1UPS[UPWARDS ARROW]NANA
Sultana et al (115)AD HippocampusPin 1ChaperoneAnti-DNPH antibody[UPWARDS ARROW]NA[DOWNWARDS ARROW]IP
 DRP-2Pyrimidine metabolism[UPWARDS ARROW]NA[DOWNWARDS ARROW]
PGM 1Glycolysis[UPWARDS ARROW]NA[DOWNWARDS ARROW]
CA IICO2 metabolism[UPWARDS ARROW][DOWNWARDS ARROW][UPWARDS ARROW]
Enolase 1Glycolysis[UPWARDS ARROW][DOWNWARDS ARROW][UPWARDS ARROW]
TPIGlycolysis[UPWARDS ARROW]=[UPWARDS ARROW]
UCHL-1UPS[UPWARDS ARROW][DOWNWARDS ARROW][UPWARDS ARROW]
Sultana et al (114)ADPin 1ChaperoneAnti-DNPH antibody[UPWARDS ARROW][DOWNWARDS ARROW][DOWNWARDS ARROW]
Sultana et al (117)ADCA IICO2 metabolismAnti-3-NTyr antibody[UPWARDS ARROW]NA[UPWARDS ARROW]IP: VDAC-1
 α-enolaseGlycolysis[UPWARDS ARROW]NA[DOWNWARDS ARROW]
GAPDHGlycolysis[UPWARDS ARROW]NA[DOWNWARDS ARROW]
ATP synthaseOxidative phosphorylation[UPWARDS ARROW]NA[UPWARDS ARROW]
VDACIon transporter[UPWARDS ARROW]NA=
Choi et al (29)ADDJ-1Antioxidant responseAnti-DNPH antibody[UPWARDS ARROW]NA[UPWARDS ARROW]Identification of oxidation sites
Newman et al (77)ADα-enolaseGlycolysisAnti-GSH antibody[UPWARDS ARROW][DOWNWARDS ARROW]NAindividual spots in 2D blots normalized with density of corresponding spots in 2D gels
 GAPDHGlycolysis[UPWARDS ARROW][DOWNWARDS ARROW]NA
Deoxyhemoglobin [UPWARDS ARROW]NANA
αB-crystallinChaperone[UPWARDS ARROW]NANA
Santpere et al (104)AD Braak stage V CAA14-3-3 protein gammaChaperone, signal transductionAnti-CEL antibody Anti-MDAL antibody[UPWARDS ARROW] PHFNANA
 14-3-3 protein zetaSignal transduction[UPWARDS ARROW] TH in AD, CAA [UPWARDS ARROW] PHF in ADNANA
Reed et al (101)AD Braak stage VPeroxiredoxin 2Antioxidant responseAnti-3-NTyr antibody[UPWARDS ARROW]NANA
 TPIGlycolysis[UPWARDS ARROW]NANA
GDHAmino acid biosynthesis[UPWARDS ARROW][DOWNWARDS ARROW]NA
Neuropolypeptide h3Phospholipid binding protein[UPWARDS ARROW]NANA
H+ transporting ATPaseOxidative phosphorylation[UPWARDS ARROW][DOWNWARDS ARROW]NA
α-enolaseGlycolysis[UPWARDS ARROW]NANA
Aldolase 1ldolase mponsites easome systemGlycolysis[UPWARDS ARROW]NANA
PGM1Glycolysis[UPWARDS ARROW]NANA
Reed et al (100)AD MCI Braak stages III, IV, V, VIHippocampus Anti-HNE antibody    
 Neuropolypeptide h3Phospholipid binding protein[UPWARDS ARROW]NANA
Carbonyl reductase 1Antioxidant response[UPWARDS ARROW]NANA
LDHGlycolysis[UPWARDS ARROW][DOWNWARDS ARROW]NA
PGKGlycolysis[UPWARDS ARROW]NANA
HSP70Chaperone[UPWARDS ARROW]NANA
ATP synthaseOxidative phosphorylation[UPWARDS ARROW][DOWNWARDS ARROW]NAIP
α-enolaseGlycolysis[UPWARDS ARROW]NANA
IPL Anti-HNE antibody    
β-actinCytoskeleton[UPWARDS ARROW]NANA
Pyruvate kinaseGlycolysis[UPWARDS ARROW][DOWNWARDS ARROW]NA
ATP synthaseOxidative phosphorylation[UPWARDS ARROW][DOWNWARDS ARROW]NA
eIF-αProtein synthesis[UPWARDS ARROW]NANA
EF-TuProtein synthesis[UPWARDS ARROW]NANA
Terni et al (119)AD Braak stages I/IIATP synthaseOxidative phosphorylationAnti-HNE antibody[UPWARDS ARROW][DOWNWARDS ARROW] complex V=Treatment with 10 mM NaBH4 for 30 min
Choi et al (26)ApoE-deficient miceGFAPCytoskeletonAnti-DNPH antibody[UPWARDS ARROW] Hippocampus = cortexNANAIn-strip DNP derivatization
 CK BBEnergy transduction[UPWARDS ARROW] Hippocampus = cortexNANA
Glucose regulated protein, Erp61Chaperone[UPWARDS ARROW] Hippocampus = cortexNANA
Chaperonin subunit 5Chaperone[UPWARDS ARROW] Hippocampus = cortexNANA
Dihydropyrimidinase-related protein 2Pyrimidine metabolism[UPWARDS ARROW] Hippocampus = cortexNANA
Mortalin, Grp70Chaperone[UPWARDS ARROW] Hippocampus = cortexNANA
Shin et al (105)Tg2576 mice bearing the APP Swedish mutationAnti-DNPH antibody Anti-DNPH antibody Anti-3NTyr antibody   
 α-enolaseGlycolysis[UPWARDS ARROW]NANA
Laminin receptor 1Neurite growth[UPWARDS ARROW]NANA
Anti-3NTyr antibody     
Atp5bOxidative phosphorylation[UPWARDS ARROW]NANA
Calpain 12Cytoskeleton remodeling processes, cell diferentiation, apoptosis, signal transduction[UPWARDS ARROW]NANA
α-enolaseGlycolysis[UPWARDS ARROW]NANA
Boyd-Kimball et al (12)Inracerebral injection of Aβ (1–42) to 3-month-old Wistar ratsNBM Anti-DNPH antibody   Sonication in re-hydration buffer on ice IP of GAPDH
 14-3-3 ζSignal transduction[UPWARDS ARROW]NANA
HSP-60Chaperone[UPWARDS ARROW]NANA
Cortex Anti-DNPH antibody    
Glutamate-ammonia ligaseAmino acid biosynthesis[UPWARDS ARROW]NANA
Tubulin β chain 15/α-tubulinCytoskeleton[UPWARDS ARROW]NANA
Hippocampus Anti-DNPH antibody    
β-synucleinRegulator of α-synuclein aggregation[UPWARDS ARROW]NANA
14-3-3 ζSignal transduction[UPWARDS ARROW]NANA
GAPDHGlycolysis[UPWARDS ARROW]NANA
Pyruvate dehydrogenaseGlycolysis-Kreb's cycle[UPWARDS ARROW]NANA
Phosphoglycerate mutase 1Glycolysis[UPWARDS ARROW]NANA
Boyd-Kimball et al (11)Intracerebral injection of Aβ (1–42) to Mongolian gerbils. Study of synaptosomal fractionsγ-actinCytoskeletonAnti-DNPH antibody[UPWARDS ARROW]NANA
 β-actinCytoskeleton[UPWARDS ARROW]NANA
GFAPCytoskeleton[UPWARDS ARROW]NANA
H+-transporting two-sector ATPaseOxidative phosphorylation[UPWARDS ARROW]NANA
Syntaxin binding protein 1Synaptic vesicle exocytosis[UPWARDS ARROW]NANA
GDHAmino acid biosynthesis[UPWARDS ARROW]NANA
Dihydropyrimidinase-related protein-2Pyrimidine metabolism[UPWARDS ARROW]NANA
EF-TuProtein synthesis[UPWARDS ARROW]NANA
Boyd-Kimball et al (13)C.elegansStrain CL 4176 (Aβ 1–42) Anti-DNPH antibody   Sonication in rehydration buffer on ice
 Medium-chain acyl-CoA dehydrogenaseFatty acid metabolism[UPWARDS ARROW]NANA
Short-chain acyl-CoA dehydrogenaseFatty acid metabolism[UPWARDS ARROW]NANA
Translation elongation factor EFProtein synthesis[UPWARDS ARROW]NANA
Malate dehydrogenaseKreb's cycle[UPWARDS ARROW]NANA
Arginine kinaseEnergy transduction[UPWARDS ARROW]NANA
RACK1 orthologAnchoring activated protein kinase C[UPWARDS ARROW]NANA
Myosin regulatory light chainCytoskeleton[UPWARDS ARROW]NANA
ActinCytoskeleton[UPWARDS ARROW]NANA
Adenosine kinasePurine biosynthesis[UPWARDS ARROW]NANA
Nematode specific proteinPresynaptic development[UPWARDS ARROW]NANA
Lipid binding proteinMetabolism[UPWARDS ARROW]NANA
TransketolaseCalvin's cycle and pentose phosphate pathway[UPWARDS ARROW]NANA
Proteasome alpha subunitUPS[UPWARDS ARROW]NANA
Proteasome beta subunitUPS[UPWARDS ARROW]NANA
Glutathionine S-transferaseAntioxidant response[UPWARDS ARROW]NANA
Strain CL 2337 Anti-DNPH antibody    
ATP synthase α chainOxidative phosphorylation[UPWARDS ARROW]NANA
Nematode-specific proteinPresynaptic development[UPWARDS ARROW]NANA
Glutamate dehydrogenaseAmino acid biosynthesis[UPWARDS ARROW]NANA
Proteasome beta subunitUPS[UPWARDS ARROW]NANA
Strain XA 1440 Anti-DNPH antibody    
20S proteasome subunit PAS-4UPS[UPWARDS ARROW]NANA
Sultana et al (116)Primary neuronal cultures treated with Aβ (1–42)GAPDHGlycolysisAnti-DNPH antibody[UPWARDS ARROW]NANASonication on ice pretreatment of the neuronal cells with D609 reduces protein oxidation
 14-3-3 zetaCell signaling[UPWARDS ARROW]NANA
Pyruvate kinaseGlycolysis[UPWARDS ARROW]NANA
MDHGlycolysis[UPWARDS ARROW]NANA

TAUOPATHIES

  1. Top of page
  2. Abstract
  3. INTRODUCTION
  4. IDENTIFICATION OF OXIDIZED AND NITRATED PROTEINS
  5. ALZHEIMER DISEASE
  6. TAUOPATHIES
  7. PARKINSON DISEASE AND RELATED α-SYNUCLEINOPATHIES
  8. HUNTINGTON DISEASE
  9. AMYOTROPHIC LATERAL SCLEROSIS
  10. COMMON AND SPECIFIC PROTEIN TARGETS IN DIFFERENT PATHOLOGIES
  11. OXIDATIVE CHANGES IN POST-MORTEM BRAIN ARE PRINCIPALLY PRIMARY
  12. PROTEIN OXIDATION IS AN EARLY EVENT IN NEURODEGENERATIVE DISEASES
  13. OXIDATIVE DAMAGE AND LOSS OF FUNCTION
  14. CLINICAL IMPLICATIONS
  15. CELLULAR LOCALIZATION OF OXIDATIVELY DAMAGED PROTEINS
  16. PITFALLS AND LIMITATIONS
  17. REFINING METHODS TO IMPROVE REDOX PROTEOMICS
  18. CONCLUDING COMMENTS
  19. ACKNOWLEDGMENTS
  20. REFERENCES
  21. Supporting Information

Studies in tauopathies including progressive supranuclear palsy (PSP), Pick disease (PiD), argyrophilic grain disease (AGD) and familial frontotemporal lobar degeneration linked to MAPT mutations (FTLD-tau) are still very limited.

Energy metabolism enzymes phosphoglycerate kinase 1 (PGK-1) and aldolase A have been shown to be oxidatively modified in frontal cortex in terminal PSP stages (65). In addition, GFAP has been identified as a major target of oxidative damage in the striatum in conventional PSP and in cases with PSP-like pathology consistent with early pre-symptomatic stages of the disease (103).

Increased oxidation of GFAP is also encountered in the amygdala in AGD (103), and in the cerebral cortex of FTLD-tau (64) and PiD (54, 74). Oxidative damage to GFAP also occurs, although to a lesser extent, in the cerebral cortex in FTLD with ubiquitin-positive, tau-negative inclusions (FTLD-U) and in FTLD associated with motor neuron disease FTLD-MND (64).

Other proteins that are targets of oxidative damage in PiD are listed as follows: vesicle-fusing ATPase, cathepsin D precursor isoforms, carbonyl reductase NADPH1 isoforms, GAPDH and HSP-7054.

Additional data of studies dealing with protein oxidative damage in selected tauopathies is shown in Table 2.

Table 2.  Tauopathies.
ReferencesDisease/stageProteinLocalization/functionDetection antibodyDegree of oxidation in relation with age-matched controlsEnzymatic activityTotal protein levelsComments
Muntanéet al (74)PiDGFAPCytoskeletonAnti-AGE, anti-CEL, anti-CML, anti-HNE, anti-MDAL antibodies[UPWARDS ARROW]NANA
Ilieva et al (54)PiDVesicle-fusing ATPaseEnergy metabolismAnti-DNPH antibody[UPWARDS ARROW]NANA
 GFAPCytoskeleton[UPWARDS ARROW]NANA
Cathepsin D precursor isoformProteolysis[UPWARDS ARROW]NANA
Carbonyl reductase NADPH1 isoformsAntioxidant response[UPWARDS ARROW]NANA
GAPDHGlycolysis[UPWARDS ARROW]NANA
HSP 70Chaperone[UPWARDS ARROW]NANA
Martínez et al (65)PSPPGK-1GlycolysisAnti-HNE antibody[UPWARDS ARROW]NA=
 Aldolasa AGlycolysis[UPWARDS ARROW]NA=
Santpere and Ferrer (103)AGD PSP, early-PSPGFAP (anti-AGE)CytoskeletonAnti-MDAL, anti-AGE, anti-CEL anti-CML antibodies[UPWARDS ARROW] Striatum in PSP [UPWARDS ARROW] Amygdala in AGDNA[UPWARDS ARROW]
Martínez et al (64)FTLD-tau FTLD-U FTLD-MNDGFAPCytoskeletonAnti-HNE antibody[UPWARDS ARROW]NA[UPWARDS ARROW]IHC Variations depending on the type of FTLD. More marked in FTLD-tau

PARKINSON DISEASE AND RELATED α-SYNUCLEINOPATHIES

  1. Top of page
  2. Abstract
  3. INTRODUCTION
  4. IDENTIFICATION OF OXIDIZED AND NITRATED PROTEINS
  5. ALZHEIMER DISEASE
  6. TAUOPATHIES
  7. PARKINSON DISEASE AND RELATED α-SYNUCLEINOPATHIES
  8. HUNTINGTON DISEASE
  9. AMYOTROPHIC LATERAL SCLEROSIS
  10. COMMON AND SPECIFIC PROTEIN TARGETS IN DIFFERENT PATHOLOGIES
  11. OXIDATIVE CHANGES IN POST-MORTEM BRAIN ARE PRINCIPALLY PRIMARY
  12. PROTEIN OXIDATION IS AN EARLY EVENT IN NEURODEGENERATIVE DISEASES
  13. OXIDATIVE DAMAGE AND LOSS OF FUNCTION
  14. CLINICAL IMPLICATIONS
  15. CELLULAR LOCALIZATION OF OXIDATIVELY DAMAGED PROTEINS
  16. PITFALLS AND LIMITATIONS
  17. REFINING METHODS TO IMPROVE REDOX PROTEOMICS
  18. CONCLUDING COMMENTS
  19. ACKNOWLEDGMENTS
  20. REFERENCES
  21. Supporting Information

Several proteins are damaged by oxidation in PD and dementia with Lewy bodies (DLB). One of them is α-synuclein, which is oxidized in the substantia nigra even at very early stages of PD (32). α-synuclein oxidative damage also occurs in the frontal cortex in PD and DLB, but also at preclinical (pre-motor) stages in which neuropathologic features (Lewy bodies) are restricted to selected nuclei of the brain stem at the time of the post-mortem study (incidental PD) (32). Other proteins, the mutations of which are causative of familial or sporadic PD, are also targets of oxidative damage in sporadic PD, such as DJ-1, which is modified by carbonylation (29). Parkin is S-nitrosated in the brain of PD (30, 126). These aspects have functional implications as oxidative stress-induced aggregation of parkin is followed by decreased parkin E3 ligase activity and impaired proteasome function (59). Down-regulation and increased oxidation of UCHL-1 has also been reported in PD (27).

In addition, increased oxidation of several glycolytic enzymes, the intensity of which increases with disease progression, has been found in PD and related diseases. Aldolase A, enolase 1 and GAPDH were oxidized, as revealed with anti-HNE antibodies, in the frontal cortex in the majority of cases of incidental PD and in all cases of PD and DLB when compared with control samples (48). Subunits of complex I have been observed to be oxidatively damaged, functionally impaired and misassembled in PD brains (56). Other proteins that are vulnerable to oxidative stress in PD are β-synuclein and SOD2 (33).

Transgenic mice overexpressing A30P mutant α-synuclein are also at risk of increased oxidative protein damage. Enolase, LDH and CA II show significantly higher carbonyl levels when compared with controls (95).

Details of studies dealing with protein targets of oxidative damage in PD and related α-synucleinopathies are found in Table 3.

Table 3.  Synucleopathies.
ReferenceDisease/stageProteinLocalization/functionDetection antibodyDegree of oxidation in relation to age-matched controlsEnzymatic activityTotal protein levelsComments
Choi et al (26)PDUCHL-1UPSAnti-DNPH antibody[UPWARDS ARROW] (10-fold)NANAIdentification of oxidation sites
Chung et al (30)PDParkinUPSAnti-S-nitro-sylated parkin antibody[UPWARDS ARROW]NA=Increase of S-nitrosylated proteins measured by the Saville method parkin S-nitrosylated by the biotin switch method
Yao et al (126)PDParkinUPSAnti-S-nitroso-parkin antibody[UPWARDS ARROW]NA=Detection of S-nitrosylated proteins by the biotin switch method
Choi et al (28)PDSOD1Antioxidant responseAnti-DNPH antibody[UPWARDS ARROW]NA[UPWARDS ARROW]Identification of oxidation sites
Dalfóet al (33)iPDβ-synucleinRegulator of α-synuclein aggregationAnti-MDAL antibody[UPWARDS ARROW]NANA
 SOD2Antioxidant response[UPWARDS ARROW]NANA
Choi et al (29)PDDJ-1Antioxidant responseAnti-DNPH antibody[UPWARDS ARROW]NA[UPWARDS ARROW]Identification of oxidation sites
Keeney et al (56)PDMitochondrial complex IElectron transport chainAnti-DNPH antibody[UPWARDS ARROW][DOWNWARDS ARROW]=Immunocapture to isolate the complete complex I in isolated mitochondria
Dalfó and Ferrer (32)iPDα-synucleinndSynapsisAnti-MDAL antibody[UPWARDS ARROW]NANAIP anti-MDAL antibody
Gómez and Ferrer (48)iPD PD DLBAldolase AGlycolysisAnti-HNE antibody[UPWARDS ARROW]NANA
 Enolase 1Glycolysis[UPWARDS ARROW]NANA
GADPHGlycolysis[UPWARDS ARROW]NANA
Poon et al (95)A30P α-synuclein transgenic miceLDHGlycolysisAnti-DNPH antibody[UPWARDS ARROW][DOWNWARDS ARROW]NA
 EnolaseGlycolysis[UPWARDS ARROW][DOWNWARDS ARROW]NA
Carbonic anhydraseCO2 metabolism[UPWARDS ARROW][DOWNWARDS ARROW]NA

HUNTINGTON DISEASE

  1. Top of page
  2. Abstract
  3. INTRODUCTION
  4. IDENTIFICATION OF OXIDIZED AND NITRATED PROTEINS
  5. ALZHEIMER DISEASE
  6. TAUOPATHIES
  7. PARKINSON DISEASE AND RELATED α-SYNUCLEINOPATHIES
  8. HUNTINGTON DISEASE
  9. AMYOTROPHIC LATERAL SCLEROSIS
  10. COMMON AND SPECIFIC PROTEIN TARGETS IN DIFFERENT PATHOLOGIES
  11. OXIDATIVE CHANGES IN POST-MORTEM BRAIN ARE PRINCIPALLY PRIMARY
  12. PROTEIN OXIDATION IS AN EARLY EVENT IN NEURODEGENERATIVE DISEASES
  13. OXIDATIVE DAMAGE AND LOSS OF FUNCTION
  14. CLINICAL IMPLICATIONS
  15. CELLULAR LOCALIZATION OF OXIDATIVELY DAMAGED PROTEINS
  16. PITFALLS AND LIMITATIONS
  17. REFINING METHODS TO IMPROVE REDOX PROTEOMICS
  18. CONCLUDING COMMENTS
  19. ACKNOWLEDGMENTS
  20. REFERENCES
  21. Supporting Information

Aldolase C, aconitase, GFAP, tubulin, peroxiredoxin 1/2/6, glutathione peroxidase and αB-crystallin were discovered as targets of oxidative modification by showing higher carbonyl levels using DNPH as a marker (107).

Increased carbonyl levels have also been shown in total homogenates of r6/2 strain transgenic HD mice, neuron-specific enolase, HSP90, aconitase, creatine kinase and VDAC have been identified as oxidized proteins (89).

AMYOTROPHIC LATERAL SCLEROSIS

  1. Top of page
  2. Abstract
  3. INTRODUCTION
  4. IDENTIFICATION OF OXIDIZED AND NITRATED PROTEINS
  5. ALZHEIMER DISEASE
  6. TAUOPATHIES
  7. PARKINSON DISEASE AND RELATED α-SYNUCLEINOPATHIES
  8. HUNTINGTON DISEASE
  9. AMYOTROPHIC LATERAL SCLEROSIS
  10. COMMON AND SPECIFIC PROTEIN TARGETS IN DIFFERENT PATHOLOGIES
  11. OXIDATIVE CHANGES IN POST-MORTEM BRAIN ARE PRINCIPALLY PRIMARY
  12. PROTEIN OXIDATION IS AN EARLY EVENT IN NEURODEGENERATIVE DISEASES
  13. OXIDATIVE DAMAGE AND LOSS OF FUNCTION
  14. CLINICAL IMPLICATIONS
  15. CELLULAR LOCALIZATION OF OXIDATIVELY DAMAGED PROTEINS
  16. PITFALLS AND LIMITATIONS
  17. REFINING METHODS TO IMPROVE REDOX PROTEOMICS
  18. CONCLUDING COMMENTS
  19. ACKNOWLEDGMENTS
  20. REFERENCES
  21. Supporting Information

Oxidative stress seems crucial in the pathogenesis of ALS (5, 53, 86). Yet practically nothing is known about protein targets of oxidative stress in ALS. GAPDH is conformationally and functionally altered in association with oxidative stress in mouse models of amyotrophic lateral sclerosis (92). Similarly, oxidative modification of SOD1, translationally controlled tumor protein, UCHL-1 and αβ-crystallin were evidenced in a mouse model of the disease (96). Transgenic mice expressing human SOD1 gene with a G93A mutation presented oxidized HSP70 and α-enolase in spinal cords, as revealed with anti-HNE antibody, and high levels of carbonyls in αβ-crystallin (88).

COMMON AND SPECIFIC PROTEIN TARGETS IN DIFFERENT PATHOLOGIES

  1. Top of page
  2. Abstract
  3. INTRODUCTION
  4. IDENTIFICATION OF OXIDIZED AND NITRATED PROTEINS
  5. ALZHEIMER DISEASE
  6. TAUOPATHIES
  7. PARKINSON DISEASE AND RELATED α-SYNUCLEINOPATHIES
  8. HUNTINGTON DISEASE
  9. AMYOTROPHIC LATERAL SCLEROSIS
  10. COMMON AND SPECIFIC PROTEIN TARGETS IN DIFFERENT PATHOLOGIES
  11. OXIDATIVE CHANGES IN POST-MORTEM BRAIN ARE PRINCIPALLY PRIMARY
  12. PROTEIN OXIDATION IS AN EARLY EVENT IN NEURODEGENERATIVE DISEASES
  13. OXIDATIVE DAMAGE AND LOSS OF FUNCTION
  14. CLINICAL IMPLICATIONS
  15. CELLULAR LOCALIZATION OF OXIDATIVELY DAMAGED PROTEINS
  16. PITFALLS AND LIMITATIONS
  17. REFINING METHODS TO IMPROVE REDOX PROTEOMICS
  18. CONCLUDING COMMENTS
  19. ACKNOWLEDGMENTS
  20. REFERENCES
  21. Supporting Information

Observations carried out by different groups have reached similar results in AD, the most studied disease, by using redox proteomics, thus giving support to the reliability of proteins vulnerable to oxidative and nitrosative stress. β-actin, β-tubulin, GFAP, α-enolase, γ-enolase, aldolase A, glutamate dehydrogenase, glutamine synthetase, ATP synthase, pyruvate kinase, UCHL-1, CK BB and Pin 1 have been identified as targets of oxidative/nitrosative damage, at least, in two different studies (see Table 1).

Also important is the evidence that several proteins linked to glycolysis and energy metabolism are targets of oxidative damage in distinct neurodegenerative diseases. Oxidative damage to aldolase A, α-enolase, LDH, UCHL-1, SOD1, DJ-1 and GAPDH have been reported in AD and PD (14, 19, 22–24, 26–29, 48, 77, 84, 100, 101, 115, 116). Increased oxidative damage to GFAP occurs in AD, tauopathies (PiD, PSP, FTLD-tau and AGD) and HD (54, 57, 65, 84, 103, 107).

Mitochondrial proteins are also vulnerable to oxidative/nitrosative stress in different conditions, although vulnerability of particular proteins appears to be disease-dependent. ATP synthase (complex V) is a target of oxidative damage in AD (84, 100, 117, 119), whereas subunits of complex I have been observed to be oxidatively damaged, functionally impaired and misassembled in PD (56).

These examples will serve to emphasize that certain proteins are damaged by oxidative stress in different pathologies whereas other proteins are selectively damaged in one degenerative disease but apparently not in another. Mitochondrial proteins are paradigms of this assumption.

Available evidence clearly indicates that only a small fraction of proteins exhibit discernible oxidative modifications, suggesting selective vulnerability. Obviously, more studies are needed to evaluate structural/functional factors shared by these proteins (if any) in order to explain this “specificity.” ROS probably act in a random fashion; however, the sensitivities and proximities of potential targets differ. The factors that can affect selectivity of oxidative damage to proteins could include the presence of a metal-binding site, molecular conformation, rate of proteolysis, relative abundance of amino acid residues susceptible to metal-catalyzed oxidation, or even protein abundance (91), among others. In this line, it is clear that modifications present in predominant proteins are easier to detect than modifications in proteins that are less abundant. Studies geared to analyze proteins that are represented at low levels in the brain will improve our understanding of selective vs. non-selective vulnerability.

However, disease-related specificities in protein vulnerability have been demonstrated as well. This is best exemplified with selective vulnerability of certain subunits of the complexes of the respiratory chain in AD and PD. ATP synthase is consistently oxidatively damaged in early stages of AD-related pathology, mild cognitive impairment with AD pathology and advanced stages of AD. In contrast, complex I is consistently altered in PD.

OXIDATIVE CHANGES IN POST-MORTEM BRAIN ARE PRINCIPALLY PRIMARY

  1. Top of page
  2. Abstract
  3. INTRODUCTION
  4. IDENTIFICATION OF OXIDIZED AND NITRATED PROTEINS
  5. ALZHEIMER DISEASE
  6. TAUOPATHIES
  7. PARKINSON DISEASE AND RELATED α-SYNUCLEINOPATHIES
  8. HUNTINGTON DISEASE
  9. AMYOTROPHIC LATERAL SCLEROSIS
  10. COMMON AND SPECIFIC PROTEIN TARGETS IN DIFFERENT PATHOLOGIES
  11. OXIDATIVE CHANGES IN POST-MORTEM BRAIN ARE PRINCIPALLY PRIMARY
  12. PROTEIN OXIDATION IS AN EARLY EVENT IN NEURODEGENERATIVE DISEASES
  13. OXIDATIVE DAMAGE AND LOSS OF FUNCTION
  14. CLINICAL IMPLICATIONS
  15. CELLULAR LOCALIZATION OF OXIDATIVELY DAMAGED PROTEINS
  16. PITFALLS AND LIMITATIONS
  17. REFINING METHODS TO IMPROVE REDOX PROTEOMICS
  18. CONCLUDING COMMENTS
  19. ACKNOWLEDGMENTS
  20. REFERENCES
  21. Supporting Information

Primary or secondary oxidative damage in post-mortem brain is difficult to ascertain as several pre-mortem factors may have produce oxidative damage. However, several complementary data support a primary origin of the observed protein modifications in post-mortem human brain. It is important to stress that targets of oxidative damage are similar in human neurodegenerative diseases, in a case-control approach, and in several animal models covering intracerebral injection of Aβ1–42 in rats and gerbils, and transgenic mice and worm models bearing human mutations of APP. Similar profiles have been reproduced in primary cortical cultures treated with Aβ1–42.

Together these data point to the likelihood that at least many proteins identified as oxidized in the post-mortem human brain are not modified as a consequence of pre-mortem agonic state but, rather, those modifications are directly linked to the degenerative process.

PROTEIN OXIDATION IS AN EARLY EVENT IN NEURODEGENERATIVE DISEASES

  1. Top of page
  2. Abstract
  3. INTRODUCTION
  4. IDENTIFICATION OF OXIDIZED AND NITRATED PROTEINS
  5. ALZHEIMER DISEASE
  6. TAUOPATHIES
  7. PARKINSON DISEASE AND RELATED α-SYNUCLEINOPATHIES
  8. HUNTINGTON DISEASE
  9. AMYOTROPHIC LATERAL SCLEROSIS
  10. COMMON AND SPECIFIC PROTEIN TARGETS IN DIFFERENT PATHOLOGIES
  11. OXIDATIVE CHANGES IN POST-MORTEM BRAIN ARE PRINCIPALLY PRIMARY
  12. PROTEIN OXIDATION IS AN EARLY EVENT IN NEURODEGENERATIVE DISEASES
  13. OXIDATIVE DAMAGE AND LOSS OF FUNCTION
  14. CLINICAL IMPLICATIONS
  15. CELLULAR LOCALIZATION OF OXIDATIVELY DAMAGED PROTEINS
  16. PITFALLS AND LIMITATIONS
  17. REFINING METHODS TO IMPROVE REDOX PROTEOMICS
  18. CONCLUDING COMMENTS
  19. ACKNOWLEDGMENTS
  20. REFERENCES
  21. Supporting Information

Pioneering studies stressed oxidative damage as an early event in AD (78, 79). In agreement with those predictions, several proteins have been identified as targets of oxidation and nitration in cases clinically manifested as mild cognitive impairment and pathologically verified as middle (IV) or early advanced (V of Braak) stages of AD-related pathology (17, 19, 20, 100, 101). More impressive, oxidative damage of ATP synthase and its associated loss of function has been observed in the entorhinal cortex in asymptomatic cases with neuropathologic AD-related pathology restricted to the entorhinal and perirhinal cortices (stage II of Braak), thus representing the earlier oxidative damage to proteins reported in AD (119).

Protein oxidative damage has also been investigated in other conditions. Protein oxidative damage was increased in brain cortex from ALS patients with lumbar debut (53). Increased oxidative damage of α-synuclein has been found in the substantia nigra at pre-clinical or pre-motor stages of PD (stages II and III of Braak) also known as incidental PD (32). Importantly, increased oxidative damage of α-synuclein, β-synuclein, SOD2, aldolase A, enolase 1 and GADPH has been shown in the cerebral cortex in incidental PD (in addition to PD and DLB) (32, 33, 48). This indicates that the cerebral cortex in PD is involved at very early stages of the disease and that oxidative damage to enzymes linked to energy metabolism and glycolysis, oxidative stress responses, and synucleins is already present at these early stages of the disease and not associated with Lewy pathology (42).

Further studies are needed to unveil oxidative damage at pre-clinical stages in tauopathies and other diseases with abnormal protein aggregates. However, recent studies have shown increased GFAP oxidative damage at pre-clinical stages of cases with PSP-like pathology (103).

OXIDATIVE DAMAGE AND LOSS OF FUNCTION

  1. Top of page
  2. Abstract
  3. INTRODUCTION
  4. IDENTIFICATION OF OXIDIZED AND NITRATED PROTEINS
  5. ALZHEIMER DISEASE
  6. TAUOPATHIES
  7. PARKINSON DISEASE AND RELATED α-SYNUCLEINOPATHIES
  8. HUNTINGTON DISEASE
  9. AMYOTROPHIC LATERAL SCLEROSIS
  10. COMMON AND SPECIFIC PROTEIN TARGETS IN DIFFERENT PATHOLOGIES
  11. OXIDATIVE CHANGES IN POST-MORTEM BRAIN ARE PRINCIPALLY PRIMARY
  12. PROTEIN OXIDATION IS AN EARLY EVENT IN NEURODEGENERATIVE DISEASES
  13. OXIDATIVE DAMAGE AND LOSS OF FUNCTION
  14. CLINICAL IMPLICATIONS
  15. CELLULAR LOCALIZATION OF OXIDATIVELY DAMAGED PROTEINS
  16. PITFALLS AND LIMITATIONS
  17. REFINING METHODS TO IMPROVE REDOX PROTEOMICS
  18. CONCLUDING COMMENTS
  19. ACKNOWLEDGMENTS
  20. REFERENCES
  21. Supporting Information

Enzymatic activity decline has been noted in AD with disease progression. Yet loss of activity may be due to reduced number of cells or specific cellular types, as well to cell redistribution, reduced amount of the enzyme or to modifications in the protein that led to protein dysfunction. All these scenarios are probably at work in advanced stages of neurodegenerative diseases. Among these possibilities, it is well known that oxidative damage of proteins has consequences in cell function (21).

Unfortunately, the majority of studies dealing with oxidized proteins in neurodegenerative diseases are not accompanied by functional studies. Perhaps historical events may account for this situation as pioneering works focused on the mere presence of increased oxidative stress and oxidative damage in aging and degenerative conditions. This was followed by the identification of DNA, RNA, lipids and proteins as targets of oxidative damage. Subsequent studies have been centered on the identification of particular proteins.

The enzymatic activities of certain oxidatively damaged proteins have been analyzed in parallel in a few studies. Increased oxidative damage accompanied by decreased activity has been shown for CK BB, enolase 1, glutamine synthetase, Pin-1, CA II, UCHL-1, α-enolase, GAPDH, GDH, H+ transporting ATPase, LDH, ATP synthase and pyruvate kinase in AD (2, 19, 77, 100, 101, 114, 115, 119). However, decreased activity can be related to lower total levels of the protein; therefore, the value of reduction due to oxidation or to the total amount of the particular protein cannot be solved in those works.

Only a few studies have included the identification of the oxidatively damaged protein, the quantification of total protein levels and the reduction of enzymatic activity (2, 114, 115, 119).

No similar data are available in PD, but oxidative damage to LDH, enolase and CA II anhydrase is associated with the corresponding decreased enzymatic activities in transgenic mice overexpressing the human A30P α-synuclein mutation (95).

CLINICAL IMPLICATIONS

  1. Top of page
  2. Abstract
  3. INTRODUCTION
  4. IDENTIFICATION OF OXIDIZED AND NITRATED PROTEINS
  5. ALZHEIMER DISEASE
  6. TAUOPATHIES
  7. PARKINSON DISEASE AND RELATED α-SYNUCLEINOPATHIES
  8. HUNTINGTON DISEASE
  9. AMYOTROPHIC LATERAL SCLEROSIS
  10. COMMON AND SPECIFIC PROTEIN TARGETS IN DIFFERENT PATHOLOGIES
  11. OXIDATIVE CHANGES IN POST-MORTEM BRAIN ARE PRINCIPALLY PRIMARY
  12. PROTEIN OXIDATION IS AN EARLY EVENT IN NEURODEGENERATIVE DISEASES
  13. OXIDATIVE DAMAGE AND LOSS OF FUNCTION
  14. CLINICAL IMPLICATIONS
  15. CELLULAR LOCALIZATION OF OXIDATIVELY DAMAGED PROTEINS
  16. PITFALLS AND LIMITATIONS
  17. REFINING METHODS TO IMPROVE REDOX PROTEOMICS
  18. CONCLUDING COMMENTS
  19. ACKNOWLEDGMENTS
  20. REFERENCES
  21. Supporting Information

Reduced oxygen uptake and impaired glycolysis have been recognized by means of neuroimaging functional studies at relatively early stages of AD manifested as mild cognitive impairment, and in advanced stages of AD. Impaired energy metabolism in the cerebral cortex has also been reported in pre-clinical stages of individuals with familial AD (8, 10, 40, 51, 69–73, 93, 94). Unfortunately, no functional neuroimaging and neuropathologic studies have been performed in the same cases. As a result, the cause of impaired energy metabolism in these cases is not known.

However, preserved protein expression levels, together with decreased enzymatic function associated with oxidative damage of relevant energy metabolism enzymes and components of the respiratory chain, have been found in a few well-documented studies (2, 114, 115, 119). These examples are particularly illuminating as they demonstrate that reduced oxygen uptake and impaired energy metabolism may be a result of oxidative damage to selected proteins rather than a consequence of neuronal loss, at least at early stages of AD-related pathology.

In the same line, mitochondrial dysfunction and impaired energy metabolism in the cerebral cortex has been demonstrated by several convergent neurologic, neuroimaging and biochemical studies in PD (42). These observations reinforce a causal link between oxidative modifications of selected proteins and functional impairment of energy metabolism in PD.

Prognostic implications of these observations are obvious because oxidative damage is subject of therapeutic intervention as oxidatively damaged molecules can be substituted by new ones whereas neuronal loss is not.

CELLULAR LOCALIZATION OF OXIDATIVELY DAMAGED PROTEINS

  1. Top of page
  2. Abstract
  3. INTRODUCTION
  4. IDENTIFICATION OF OXIDIZED AND NITRATED PROTEINS
  5. ALZHEIMER DISEASE
  6. TAUOPATHIES
  7. PARKINSON DISEASE AND RELATED α-SYNUCLEINOPATHIES
  8. HUNTINGTON DISEASE
  9. AMYOTROPHIC LATERAL SCLEROSIS
  10. COMMON AND SPECIFIC PROTEIN TARGETS IN DIFFERENT PATHOLOGIES
  11. OXIDATIVE CHANGES IN POST-MORTEM BRAIN ARE PRINCIPALLY PRIMARY
  12. PROTEIN OXIDATION IS AN EARLY EVENT IN NEURODEGENERATIVE DISEASES
  13. OXIDATIVE DAMAGE AND LOSS OF FUNCTION
  14. CLINICAL IMPLICATIONS
  15. CELLULAR LOCALIZATION OF OXIDATIVELY DAMAGED PROTEINS
  16. PITFALLS AND LIMITATIONS
  17. REFINING METHODS TO IMPROVE REDOX PROTEOMICS
  18. CONCLUDING COMMENTS
  19. ACKNOWLEDGMENTS
  20. REFERENCES
  21. Supporting Information

Seminal immunohistochemical studies carried out several years ago showed that neurons and glial cells were putative sources of oxidative stress in the nervous system as they were labeled with antibodies that recognize oxidative adducts. Moreover, certain oxidative stress responses are particularly robust in astroglia.

In contrast with these observations, it is assumed that oxidatively damaged proteins, as detected by redox proteomics, are predominantly neuronal proteins. In fact there is little evidence that a particular damaged protein is neuronal or glial, unless the localization of the protein is known in advance. It is logical to interpret that oxidatively damaged neurofilaments are localized in neurons, whereas oxidized glial fibrillary protein is localized in astrocytes. In some examples, HNE adducts co-localize with GFAP in astrocytes, as revealed by double-labeling immunofluorescence and confocal microscopy, at the time that HNE-modified GFAP is identified by bidimensional gel electrophoresis, Western blotting, in-gel digestion and mass spectrometry (64).

It is clear that further studies are needed to elucidate the localization of damaged molecules in neurons and glial cells to understand the implications of the abnormalities in definite cellular types. This is not only valid considering neurons vs. glial cells, but also among different neuronal types. The fact that similar molecules are oxidatively damaged in variegated degenerative diseases does not prove that the same neurons are affected in the different conditions.

PITFALLS AND LIMITATIONS

  1. Top of page
  2. Abstract
  3. INTRODUCTION
  4. IDENTIFICATION OF OXIDIZED AND NITRATED PROTEINS
  5. ALZHEIMER DISEASE
  6. TAUOPATHIES
  7. PARKINSON DISEASE AND RELATED α-SYNUCLEINOPATHIES
  8. HUNTINGTON DISEASE
  9. AMYOTROPHIC LATERAL SCLEROSIS
  10. COMMON AND SPECIFIC PROTEIN TARGETS IN DIFFERENT PATHOLOGIES
  11. OXIDATIVE CHANGES IN POST-MORTEM BRAIN ARE PRINCIPALLY PRIMARY
  12. PROTEIN OXIDATION IS AN EARLY EVENT IN NEURODEGENERATIVE DISEASES
  13. OXIDATIVE DAMAGE AND LOSS OF FUNCTION
  14. CLINICAL IMPLICATIONS
  15. CELLULAR LOCALIZATION OF OXIDATIVELY DAMAGED PROTEINS
  16. PITFALLS AND LIMITATIONS
  17. REFINING METHODS TO IMPROVE REDOX PROTEOMICS
  18. CONCLUDING COMMENTS
  19. ACKNOWLEDGMENTS
  20. REFERENCES
  21. Supporting Information

With no doubt, redox proteomics is a useful tool for detecting damaged proteins resulting from oxidative/nitrosative damage. Yet available studies have also detected limitations and pitfalls.

First of all, information about post-mortem delay is important as certain proteins might be affected by oxidation/nitration with post-mortem delay. On the other hand, degradation of proteins with post-mortem delay may minimize the abundance of oxidized proteins due to non-specific post-mortem degradation. Several available studies take into consideration this aspect whereas this information is lacking in many others. A caveat derived from this observation is that identification of oxidized/nitrated proteins in human neurodegenerative diseases is feasible provided that tissue samples are examined within a time not surpassing thresholds of protein degradation and vulnerability to oxidative damage which are variable from one protein to another.

A second point is the overrepresentation of most abundant proteins whereas oxidative damage of minority proteins is probably underdetected. This may lead to an oversimplification of damaged metabolic pathways and, therefore, to the putative neglect of damaged crucial components.

A third point is the lack of information regarding cellular types involved, including particular neuronal types in different neurodegenerative disorders. We barely understand the reasons for selective cell vulnerability in general terms, and this shortage may also be applied to selective vulnerability of individual molecules in particular cell types.

Another source of possible confusion is based on the convergence of abnormalities in particular metabolic pathways that may obscure the real impact of oxidative damage in determined cellular functions. As an example, abnormal mitochondrial function in AD may be the result of several components, including increased mitochondrial DNA deletions, abnormal fusion and fission of mitochondria, and decreased expression of certain complexes of the respiratory chain, such as complex IV (45, 52, 68, 87, 125). Mitochondrial alterations result in increased oxidative stress. Oxidative damage to ATP synthase (complex V), VDAC, ubiquinol–cytochrome c reductase complex core protein I, H+ transporting ATPase and Atp5b (11, 13, 58, 84, 100, 105, 117, 119) may, in turn, increase mitochondrial dysfunction.

Similar considerations can be applied to the cerebral cortex in PD (42, 76).

REFINING METHODS TO IMPROVE REDOX PROTEOMICS

  1. Top of page
  2. Abstract
  3. INTRODUCTION
  4. IDENTIFICATION OF OXIDIZED AND NITRATED PROTEINS
  5. ALZHEIMER DISEASE
  6. TAUOPATHIES
  7. PARKINSON DISEASE AND RELATED α-SYNUCLEINOPATHIES
  8. HUNTINGTON DISEASE
  9. AMYOTROPHIC LATERAL SCLEROSIS
  10. COMMON AND SPECIFIC PROTEIN TARGETS IN DIFFERENT PATHOLOGIES
  11. OXIDATIVE CHANGES IN POST-MORTEM BRAIN ARE PRINCIPALLY PRIMARY
  12. PROTEIN OXIDATION IS AN EARLY EVENT IN NEURODEGENERATIVE DISEASES
  13. OXIDATIVE DAMAGE AND LOSS OF FUNCTION
  14. CLINICAL IMPLICATIONS
  15. CELLULAR LOCALIZATION OF OXIDATIVELY DAMAGED PROTEINS
  16. PITFALLS AND LIMITATIONS
  17. REFINING METHODS TO IMPROVE REDOX PROTEOMICS
  18. CONCLUDING COMMENTS
  19. ACKNOWLEDGMENTS
  20. REFERENCES
  21. Supporting Information

The large majority of studies in human brain and animal models have been carried out by analyzing total homogenates of a particular region. This is with no doubt an adequate approach although damage of minority proteins can be easily underrepresented. The study of cellular fractions may improve the resolution of the study by increasing the total amount of a particular protein. In this line, sarkosyl-insoluble fractions have been used to recover proteins in paired-helical filament-enriched fractions (104), mitochondria-enriched fractions to recover proteins principally related with mitochondria (30, 48), or synaptosomal-enriched fractions to reveal abnormalities of synapsis-related proteins (11).

Another important point is the use of different protocols and buffers to increase the capture of different proteins. The combination of different buffers to grind or to focus samples is a strategy to improve the reproducibility at the acidic or alkaline extremes of the electrophoresis gel and, likewise, to solubilize different proteins receiving a better number of spots and resolution (9). This observation, originally applied to general bidimensional gel electrophoresis methodology, may be of considerable interest in redox proteomics to optimize spots detected as oxidized proteins. We should also take into account that membrane proteins are difficult to detect by current bidimensional gel electrophoresis, thus probably accounting for the low numbers of membrane proteins identified as targets of oxidation.

Finally, detection of specific residues of oxidative modifications may increase understanding of specific oxidation sites and their relevance to protein function. However, this approach is time-consuming and a combination of different methods is needed. By using MALDI-TOF/MS and HPLC-ESI/MS/MS techniques, oxidation sites have been identified only in UCH-L1, SOD1 and DJ-1 in AD and PD as yet (27, 28).

CONCLUDING COMMENTS

  1. Top of page
  2. Abstract
  3. INTRODUCTION
  4. IDENTIFICATION OF OXIDIZED AND NITRATED PROTEINS
  5. ALZHEIMER DISEASE
  6. TAUOPATHIES
  7. PARKINSON DISEASE AND RELATED α-SYNUCLEINOPATHIES
  8. HUNTINGTON DISEASE
  9. AMYOTROPHIC LATERAL SCLEROSIS
  10. COMMON AND SPECIFIC PROTEIN TARGETS IN DIFFERENT PATHOLOGIES
  11. OXIDATIVE CHANGES IN POST-MORTEM BRAIN ARE PRINCIPALLY PRIMARY
  12. PROTEIN OXIDATION IS AN EARLY EVENT IN NEURODEGENERATIVE DISEASES
  13. OXIDATIVE DAMAGE AND LOSS OF FUNCTION
  14. CLINICAL IMPLICATIONS
  15. CELLULAR LOCALIZATION OF OXIDATIVELY DAMAGED PROTEINS
  16. PITFALLS AND LIMITATIONS
  17. REFINING METHODS TO IMPROVE REDOX PROTEOMICS
  18. CONCLUDING COMMENTS
  19. ACKNOWLEDGMENTS
  20. REFERENCES
  21. Supporting Information

Neurodegenerative diseases with abnormal protein aggregates are associated with modifications of solubility, aggregation and fibril formation of selected proteins. Mutant proteins resulting from DNA mutations are causative in familial and certain sporadic settings. More commonly, post-translational modifications of proteins are involved in the majority of sporadic cases. Understanding the mechanisms involved in such modifications is crucial from a mechanistic perspective, but it is also essential for the delineation of therapeutic strategies. Oxidative stress plays a functional role in physiologic conditions as it switches on vital cellular responses. Yet imbalance between oxidative stress sources and antioxidant responses may cause a net flux of oxidative damage to DNA, RNA, carbohydrates, lipids and proteins, and, in most cases, concomitant loss of function. Oxidative stress increases with age largely because of progressive mitochondrial dysfunction and impairment or loss of cellular repair mechanisms. In addition, neurodegenerative diseases are associated with higher levels of oxidative damage and higher levels of direct and indirect protein modifications resulting from increased oxidation, nitrosation and nitration when compared with those occurring in age-matched individuals with no diseases of the nervous system.

Vulnerable proteins can be grouped in defined metabolic pathways covering glycolysis and energy metabolism, mitochondrial proteins, cytoskeleton, chaperones, and members of the ubiquitin–proteasome system, among many others. Some proteins are affected in different degenerative diseases whereas others appear to be disease-specific. Importantly, many damaged proteins in human neurodegenerative diseases are also damaged in experimental models, transgenic mice and worms, and cell culture paradigms. These findings indicate that oxidative stress observed in post-mortem brains is a primary event linked to degeneration rather than a secondary effect resulting from pre-mortem agonic states.

Since oxidative damage may result in impaired function, protein oxidative damage may have important consequences on the nervous system thus resulting in abnormal glycolysis and energy metabolism, abnormal responses to protein folding and oxidative stress responses, cytoskeletal abnormalities, and impaired protein degradation, in addition to damage to relevant proteins as α-synuclein in PD and related α-synucleinopathies. Some of these abnormalities are reflected in vivo by using sophisticated metabolic and neuroimaging methods. Thus, abnormal energy metabolism has been observed in the cerebral cortex not only in AD but also in patients with mild cognitive impairment and in patients with PD in whom impaired metabolism cannot be ascribed to neuron loss. Therefore, it is reasonable to think that part of the metabolic disturbances observed at early stages of degenerative processes are related to oxidative damage of selected proteins rather than to neuron loss. However, much work has to be done as the majority of redox proteomics studies are not accompanied by functional analysis of oxidatively damaged proteins.

Available information points to the fact that vital metabolic pathways are hampered by protein oxidative damage in several human degenerative diseases at very early stages of the disease. A better understanding of proteins susceptible to oxidation and nitration may serve to define damaged metabolic networks at early stages of disease and to procure therapeutic interventions to attenuate disease progression.

ACKNOWLEDGMENTS

  1. Top of page
  2. Abstract
  3. INTRODUCTION
  4. IDENTIFICATION OF OXIDIZED AND NITRATED PROTEINS
  5. ALZHEIMER DISEASE
  6. TAUOPATHIES
  7. PARKINSON DISEASE AND RELATED α-SYNUCLEINOPATHIES
  8. HUNTINGTON DISEASE
  9. AMYOTROPHIC LATERAL SCLEROSIS
  10. COMMON AND SPECIFIC PROTEIN TARGETS IN DIFFERENT PATHOLOGIES
  11. OXIDATIVE CHANGES IN POST-MORTEM BRAIN ARE PRINCIPALLY PRIMARY
  12. PROTEIN OXIDATION IS AN EARLY EVENT IN NEURODEGENERATIVE DISEASES
  13. OXIDATIVE DAMAGE AND LOSS OF FUNCTION
  14. CLINICAL IMPLICATIONS
  15. CELLULAR LOCALIZATION OF OXIDATIVELY DAMAGED PROTEINS
  16. PITFALLS AND LIMITATIONS
  17. REFINING METHODS TO IMPROVE REDOX PROTEOMICS
  18. CONCLUDING COMMENTS
  19. ACKNOWLEDGMENTS
  20. REFERENCES
  21. Supporting Information

Work carried out at the Institute of Neuropathology was partially funded by grants from the Spanish Ministry of Health, Instituto de Salud Carlos III PI080582, and supported by the European Commission under the Sixth Framework Programme BrainNet Europe II, LSHM-CT-2004-503039 and INDABIP FP6-2005-LIFESCIHEALTH-7 Molecular Diagnostics. Work carried out at the Department of Experimental Medicine was supported in part by I+D grants from the Spanish Ministry of Education and Science (BFU2006-14495/BFI and AGL2006-12433), the Spanish Ministry of Health (ISCIII, Red de Envejecimiento y Fragilidad, RD06/0013/0012, PI081843), the Autonomous Government of Catalonia (2005SGR00101), “La Caixa” Foundation and COST B-35 Action.

We thank Odena MA and Oliveira E from the Proteomics platform, Science Park, University of Barcelona, for support.

Thanks to T. Yohannan for editorial help. There is no conflict of interest including any financial, personal or other relationships with other people or organizations within the three years from the beginning of the work.

REFERENCES

  1. Top of page
  2. Abstract
  3. INTRODUCTION
  4. IDENTIFICATION OF OXIDIZED AND NITRATED PROTEINS
  5. ALZHEIMER DISEASE
  6. TAUOPATHIES
  7. PARKINSON DISEASE AND RELATED α-SYNUCLEINOPATHIES
  8. HUNTINGTON DISEASE
  9. AMYOTROPHIC LATERAL SCLEROSIS
  10. COMMON AND SPECIFIC PROTEIN TARGETS IN DIFFERENT PATHOLOGIES
  11. OXIDATIVE CHANGES IN POST-MORTEM BRAIN ARE PRINCIPALLY PRIMARY
  12. PROTEIN OXIDATION IS AN EARLY EVENT IN NEURODEGENERATIVE DISEASES
  13. OXIDATIVE DAMAGE AND LOSS OF FUNCTION
  14. CLINICAL IMPLICATIONS
  15. CELLULAR LOCALIZATION OF OXIDATIVELY DAMAGED PROTEINS
  16. PITFALLS AND LIMITATIONS
  17. REFINING METHODS TO IMPROVE REDOX PROTEOMICS
  18. CONCLUDING COMMENTS
  19. ACKNOWLEDGMENTS
  20. REFERENCES
  21. Supporting Information
  • 1
    Adams S, Green P, Claxton R, Simcox S, Williams MV, Walsh K, Leewenburgh C (2001) Reactive carbonyl formation by oxidative and non-oxidative pathways. Front Biosci 6:A17A24.
  • 2
    Aksenov M, Aksenova M, Butterfield DA, Markesbery WR (2000) Oxidative modification of creatine kinase BB in Alzheimer's disease brain. J Neurochem 74:25202527.
  • 3
    Aksenov MY, Atiksenova MV, Butterfield DA, Geddes JW, Markesbery WR (2001) Protein oxidation in the brain in Alzheimer's disease. Neuroscience 103:373383.
  • 4
    Aruoma OI, Kaur H, Halliwell B (1991) Oxygen free radicals and human diseases. J R Soc Health 111:172177.
  • 5
    Barber SC, Mead RJ, Shaw PJ (2006) Oxidative stress in ALS: a mechanism of neurodegeneration and a therapeutic target. Biochim Biophys Acta 1762:10511067.
  • 6
    Barja G (2007) Mitochondrial oxygen consumption and reactive oxygen species production are independently modulated: implications for aging studies. Rejuvenation Res 10:215224.
  • 7
    Barnham KJ, Masters CL, Bush AI (2004) Neurodegenerative diseases and oxidative stress. Nat Rev Drug Discov 3:205214.
  • 8
    Bigl M, Brückner MK, Arendt T, Bigl V, Eschrich K (1999) Activities of key glycolytic enzymes in the brains of patients with Alzheimer's disease. J Neural Transm 106:499511.
  • 9
    Bland AM, D'Eugenio LR, Dugan MA, Janech MG, Almeida JS, Zile MR, Arthur JM (2006) Comparison of variability associated with sample preparation in two-dimensional gel electrophoresis of cardiac tissue. J Biomol Tech 17:195199.
  • 10
    Blass JP (2002) Alzheimer's disease and Alzheimer's dementia: distinct but overlapping entities. Neurobiol Aging 23:10771084.
  • 11
    Boyd-Kimball D, Castegna A, Sultana R, Poon HF, Petroze R, Lynn BC et al (2005) Proteomic identification of proteins oxidized by Aβ(1–42) in synaptosomes: implications for Alzheimer's disease. Brain Res 1044:206215.
  • 12
    Boyd-Kimball D, Sultana R, Poon HF, Lynn BC, Casamenti F, Pepeu G et al (2005) Proteomic identification of proteins specifically oxidized by intracerebral injection of amyloid β-peptide (1–42) into rat brain: implications for Alzheimer's disease. Neuroscience 132:313324.
  • 13
    Boyd-Kimball D, Poon HF, Lynn BC, Cai J, Pierce WM Jr, Klein JB et al (2006) Proteomic identification of proteins specifically oxidized in Caenorhabditis elegans expressing human Aβ(1–42): implications for Alzheimer's disease. Neurobiol Aging 27:12391249.
  • 14
    Butterfield DA, Kanski J (2001) Brain protein oxidation in age-related neurodegenerative disorders that are associated with aggregated proteins. Mech Ageing Dev 122:945962.
  • 15
    Butterfield DA, Stadtman ER (1997) Protein oxidation processes in aging brain. In: Advances in Cell Aging and Gerontology, Vol. 2. PSTimiras, EEBittar (eds), pp. 161191. JAI Press, Greenwich.
  • 16
    Butterfield DA, Sultana R (2008) Redox proteomics: understanding oxidative stress in the progression of age-related neurodegenerative disorders. Expert Rev Proteomics 5:157160.
  • 17
    Butterfield DA, Abdul HM, Newman S, Reed T (2006) Redox proteomics in some age-related neurodegenerative disorders or models thereof. NeuroRx 3:344357.
  • 18
    Butterfield DA, Gnjec A, Poon HF, Castegna A, Pierce WM, Klein JB, Martins RN (2006) Redox proteomics identification of oxidatively modified brain proteins in inherited Alzheimer's disease: an initial assessment. J Alzheimers Dis 10:391397.
  • 19
    Butterfield DA, Poon HF, St Clair D, Keller JN, Pierce WM, Klein JB, Markesbery WR (2006) Redox proteomics identification of oxidatively modified hippocampal proteins in mild cognitive impairment: insights into the development of Alzheimer's disease. Neurobiol Dis 22:223232.
  • 20
    Butterfield DA, Sultana R, Poon HF (2006) Redox proteomics: a new approach to investigate oxidative stress in Alzheimer's disease. In: Redox Proteomics: From Protein Modifications to Cellular Dysfunction and Diseases. IDalle-Donne, AScaloni, DAButterfield (eds), pp. 563603. John Wiley & Sons: Hoboken, NJ.
  • 21
    Cabiscol E, Ros J (2006) Oxidative damage to proteins: structural modifications and consequences in cell function. In: Redox Proteomics: From Protein Modifications to Cellular Dysfunction and Diseases. IDalle-Donne, AScaloni, DAButterfield (eds), pp. 399471. John Wiley & Sons: Hoboken, NJ.
  • 22
    Castegna A, Aksenov M, Aksenova M, Thongboonkerd V, Klein JB, Pierce WM et al (2002) Proteomic identification of oxidatively modified proteins in Alzheimer's disease brain. Part I: creatine kinase BB, glutamine synthase, and ubiquitin carboxy-terminal hydrolase L-1. Free Radic Biol Med 33:562571.
  • 23
    Castegna A, Aksenov M, Thongboonkerd V, Klein JB, Pierce WM, Booze R et al (2002) Proteomic identification of oxidatively modified proteins in Alzheimer's disease brain. Part II: dihydropyrimidinase-related protein 2, α-enolase and heat shock cognate 71. J Neurochem 82:15241532.
  • 24
    Castegna A, Thongboonkerd V, Klein JB, Lynn B, Markesbery WR, Butterfield DA (2003) Proteomic identification of nitrated proteins in Alzheimer's disease brain. J Neurochem 85:13941401.
  • 25
    Chinta SJ, Andersen JK (2008) Redox imbalance in Parkinson's disease. Biochem Biophys Acta 1780:13621367.
  • 26
    Choi J, Forster MJ, McDonald SR, Weintraub ST, Carroll CA, Gracy RW (2004) Proteomic identification of specific oxidized proteins in ApoE-knockout mice: relevance to Alzheimer's disease. Free Radic Biol Med 36:11551162.
  • 27
    Choi J, Levey AI, Weintraub ST, Rees HD, Gearing M, Chin LS, Li L (2004) Oxidative modifications and down-regulation of ubiquitin carboxyl-terminal hydrolase L1 associated with idiopathic Parkinson's and Alzheimer's diseases. J Biol Chem 279:1325613264.
  • 28
    Choi J, Rees HD, Weintraub ST, Levey AI, Chin LS, Li L (2005) Oxidative modifications and aggregation of Cu,Zn-superoxide dismutase associated with Alzheimer and Parkinson diseases. J Biol Chem 280:1164811655.
  • 29
    Choi J, Sullards MC, Olzmann JA, Rees HD, Weintraub ST, Bostwick DE et al (2006) Oxidative damage of DJ-1 is linked to sporadic Parkinson and Alzheimer diseases. J Biol Chem 281:1081610824.
  • 30
    Chung KKK, Thomas B, Li X, Pletnikova O, Troncoso JC, Marsh L et al (2004) S-nitrosylation of parkin regulates ubiquitination and compromises parkin's protective function. Science 304:13281331.
  • 31
    Cosgrove JP, Church DF, Pryor WA (1987) The kinetics of the autoxidation of polyunsaturated fatty acids. Lipids 22:299304.
  • 32
    Dalfó E, Ferrer I (2008) Early α-synuclein lipoxidation in neocortex in Lewy body diseases. Neurobiol Aging 29:408417.
  • 33
    Dalfó E, Portero-Otín M, Ayala V, Martínez A, Pamplona R, Ferrer I (2005) Evidence of oxidative stress in the neocortex in incidental Lewy body disease. J Neuropathol Exp Neurol 64:816830.
  • 34
    Dalle-Donne I, Scaloni A, Giustarini D, Cavarra E, Tell G, Lungarella G et al (2005) Proteins as biomarkers of oxidative/nitrosative stress in diseases: the contribution of redox proteomics. Mass Spectrom Rev 24:5599.
  • 35
    Dalle-Donne I, Aldini G, Carini M, Colombo R, Rossi R, Milzani A (2006) Protein carbonylation, cellular dysfunction, and disease progression. J Cell Mol Med 10:389406.
  • 36
    Dalle-Donne I, Rossi R, Ceciliani F, Giustarini D, Colombo R, Milzani A (2006) Proteins as sensitive biomarkers of human conditions associated with oxidative stress. In: Redox Proteomics: From Protein Modifications to Cellular Dysfunction and Diseases. IDalle-Donne, AScaloni, DAButterfield (eds), pp. 487525. John Wiley & Sons: Hoboken, NJ.
  • 37
    Dalle-Donne I, Scaloni A, Butterfield DA (eds) (2006) Redox Proteomics: From Protein Modifications to Cellular Dysfunction and Diseases. John Wiley & Sons: Hoboken, NJ.
  • 38
    Daneshvar B, Frandsen H, Autrup H, Dragsted LO (1997) γ-Glutamyl semialdehyde and 2-amino-adipic semialdehyde: biomarkers of oxidative damage to proteins. Biomarkers 2:117123.
  • 39
    Danielson SR, Andersen JK (2008) Oxidative and nitrative protein modifications in Parkinson's disease. Free Radic Biol Med 44:17871794.
  • 40
    De Leon MJ, Convit A, Wolf OT, Tarshish CY, DeSanti S, Rusinek H et al (2001) Prediction of cognitive declinein normal elderly subjects with 2-[18F]fluorodo-2-deoxy-D-glucosee/positron-emission tomography (FDG/PET). Proc Natl Acad Sci USA 98:1096610971.
  • 41
    Ding Q, Dimayuga E, Keler JN (2007) Oxidative damage, protein synthesis, and protein degradation in Alzheimer's disease. Curr Alzheimer Res 4:7379.
  • 42
    Ferrer I (2009) Early involvement of the cerebral cortex in Parkinson's disease: convergence of multiple metabolic defects. Progr Neurobiol 88:89103.
  • 43
    Ferrer I, Martinez A, Boluda S, Parchi P, Barrachina M (2008) Brain banks: benefits, limitations and cautions concerning the use of post-mortem brain tissue for molecular studies. Cell Tissue Bank 9:181194.
  • 44
    Gibson GE, Huang HM (2005) Oxidative stress in Alzheimer's disease. Neurobiol Aging 26:575578.
  • 45
    Gibson GE, Karuppagounder SS, Shi Q (2008) Oxidant-induced changes in mitochondria and calcium dynamics in the pathophysiology of Alzheimer's disease. Ann N Y Acad Sci 1147:221232.
  • 46
    Gibson GE, Ratan RR, Beal MF (2008) Mitochondria and oxidatiuve stress in neurodegenerative disorders. Preface. Ann NY Acad Sci 1147:xixii.
  • 47
    Gilca M, Stoian I, Atanasiu V, Virgolici B (2007) The oxidative hypothesis of senescence. J Postgrad Med 53:207213.
  • 48
    Gómez A, Ferrer I (2009) Increased oxidation of certain glycolysis and energy metabolism enzymes in the frontal cortex in Lewy body diseases. J Neurosci Res 87:10021013.
  • 49
    Halliwell B (1991) Reactive oxygen species in living systems: source, biochemistry, and role in human disease. Am J Med 91:14S22S.
  • 50
    Halliwell B, Gutteridge JMC (2007) Free Radicals in Biology and Medicine. Oxford University Press: New York.
  • 51
    Herholz K, Carter SF, Jones M (2007) Positron emission tomography imaging in dementia. Br J Radiol 80(Spec. No. 2):S160S167.
  • 52
    Hirai K, Aliev G, Nunomura A, Fujioka H, Rusell RL, Atwood CS et al (2001) Mitochondrial abnormalities in Alzheimer's disease. J Neurosci 21:30173023.
  • 53
    Ilieva EV, Ayala V, Jové M, Dalfó E, Cacabelos D, Povedano M et al (2007) Oxidative and endoplasmic reticulum stress interplay in sporadic amyotrophic lateral sclerosis. Brain 130:31113123.
  • 54
    Ilieva EV, Naudí A, Kichev A, Ferrer I, Pamplona R, Portero-Otín M (2009) Loss of the Stress Transducers Nrf2 and Grp78/BiP in Pick's Disease. (In press).
  • 55
    Ischiropoulos H, Al-Medi AB (1995) Peroxynitrite-mediated oxidative protein modifications. FEBS Lett 364:279282.
  • 56
    Keeney PM, Xie J, Capaldi RA, Bennett JP (2006) Parkinson's disease brain mitochondrial complex I has oxidatively damaged subunits and is functionally impaired and misassembled. J Neurosci 26:52565264.
  • 57
    Korolainen MA, Auriola S, Nyman TA, Alafuzoff I, Pirttilä T (2005) Proteomic analysis of glial fibrillary acidic protein in Alzheimer's disease and aging brain. Neurobiol Dis 20:858870.
  • 58
    Korolainen MA, Goldsteins G, Nyman TA, Alafuzoff I, Koistinaho J, Pirttilä T (2006) Oxidative modification of proteins in the frontal cortex of Alzheimer's disease brain. Neurobiol Aging 27:4253.
  • 59
    LaVoie MJ, Cortese GP, Ostaszewski BL, Schlossmacher MG (2007) The effects of oxidative stress on parkin and other E3 ligases. J Neurochem 103:23542368.
  • 60
    Levine RL, Stadtman ER (2001) Oxidative modifications of proteins during aging. Exp Gerontol 36:14951502.
  • 61
    Lowell MA, Markesbery WB (2007) Oxidative DNA damage in mild cognitive impairment and late-stage Alzheimer's disease. Nucl Acid Res 35:74977504.
  • 62
    Mancuso C, Scapagini G, Currò D, Giuffrida Stella AM, De Marco C, Butterfield DA, Calabrese V (2007) Mitochondrial dysfunction, free radical generation and cellular stress response in neurodegenerative disorders. Front Biosci 12:11071123.
  • 63
    Markesbery WR, Lowell MA (2007) Damage to lipids, proteins, DNA and RNA in mild cognitive impairment. Arch Neurol 64:954956.
  • 64
    Martínez A, Carmona M, Portero-Otin M, Naudí A, Pamplona R, Ferrer I (2008) Type-dependent oxidative damage in frontotemporal lobar degeneration: cortical astrocytes are targets of oxidative damage. J Neuropathol Exp Neurol 67:11221136.
  • 65
    Martínez A, Dalfó E, Muntané G, Ferrer I (2008) Glycolitic enzymes are targets of oxidation in aged human frontal cortex and oxidative damage of these proteins is increased in progressive supranuclear palsy. J Neural Transm 115:5966.
  • 66
    Mikkelsen RB, Wardman P (2003) Biological, chemistry of reactive oxygen and nitrogen and radiation-induced signal transduction mechanisms. Oncogene 22:57345754.
  • 67
    Moreira PI, Smith MA, Zhu X, Nunomura A, Castellani RJ, Perry G (2005) Oxidative stress and neurodegeneration. Ann N Y Acad Sci 1043:545552.
  • 68
    Moreira PI, Santos MS, Oliveira CR (2007) Alzheimer's disease: a lesson from mitochondrial dysfunction. Antioxid Redox Signal 9:16211630.
  • 69
    Mosconi L (2005) Brain glucose metabolism in the early and specific diagnosis of Alzheimer's disease. Eur J Nucl Med 32:486510.
  • 70
    Mosconi L, De Santi S, Li J, Tsui WH, Li Y, Boppana M et al (2006) Hippocampal metabolism predicts cognitive decline from normal aging. Neurobiol Aging 29:676692.
  • 71
    Mosconi L, Sorbi S, De Leon MJ, Li Y, Nacmias B, Myoung PS et al (2006) Hypometabolism exceeds atrophy in presymptomatic early-onset familial Alzheimer's disease. J Nucl Med 47:17781786.
  • 72
    Mosconi L, Pupi A, De Leon MJ (2008) Brain glucose metabolism and oxidative stress in preclinical Alzheimer's disease. Ann N Y Acad Sci 1147:180195.
  • 73
    Mosconi L, Tsui WH, Herholz K, Pupi A, Drzezga A, Lucignani G et al (2008) Multicenter standardized 18F-FDG PET diagnosis of mild cognitive impairment, Alzheimer's disease, and other dementias. J Nucl Med 49:390398.
  • 74
    Muntané G, Dalfó E, Martínez A, Rey MJ, Avila J, Pérez M et al (2006) Glial fibrillary acidic protein is a major target of glycoxidative and lipoxidative damage in Pick's disease. J Neurochem 99:177185.
  • 75
    Nakabeppu Y, Tsuchimoto D, Yamaguchi H, Sakumi K (2007) Oxidative damage in nucleic acids and Parkinson's disease. J Neurosci Res 85:919934.
  • 76
    Navarro A, Boveris A, Bández MJ, Sanchez-Pinto MJ, Gómez C, Muntane G, Ferrer I (2009) Human brain cortex: mitochondrial oxidative damage and adaptative response in Parkinson disease and dementia with Lewy bodies. Free Radic Biol Med 46:15741580.
  • 77
    Newman SF, Sultana R, Perluigi M, Coccia R, Cai J, Pierce WM et al (2007) An increase in S-glutathionylated proteins in the Alzheimer's disease inferior parietal lobule, a proteomics approach. J Neurosci Res 85:15061514.
  • 78
    Nunomura A, Perry G, Aliev G, Hirai K, Takeda A, Balraj EK et al (2001) Oxidative damage is the earliest event in Alzheimer disease. J Neuropathol Exp Neurol 60:759767.
  • 79
    Nunomura A, Castellani RJ, Zhu X, Moreira PI, Perry G, Smith MA (2006) Involvement of oxidative stress in Alzheimer disease. J Neuropathol Exp Neurol 65:631641.
  • 80
    Nunomura A, Moreira PI, Takeda A, Smith MA, Perry G (2007) Oxidative RNA damage and neurodegeneration. Curr Med Chem 14:29682975.
  • 81
    Onyango IG, Khan SM (2006) Oxidative stress, mitochondrial dysfunction, and stress signalling in Alzheimer's disease. Curr Alzheimer Res 3:339349.
  • 82
    Pamplona R (2008) Membrane phospholipids, lipoxidative damage and molecular integrity: a causal role in aging and longevity. Biochim Biophys Acta 1777:12491262.
  • 83
    Pamplona R, Barja G (2007) Highly resistant macromolecular components and low rate of generation of endogenous damage: two key traits of longevity. Ageing Res Rev 6:189210.
  • 84
    Pamplona R, Dalfó E, Ayala V, Bellmunt MJ, Prat J, Ferrer I, Portero-Otín M (2005) Proteins in human brain cortex are modified by oxidation, glycoxidation, and lipoxidation. Effects of Alzheimer disease and identification of lipoxidation targets. J Biol Chem 280:2152221530.
  • 85
    Pamplona R, Ilieva E, Ayala V, Bellmunt MJ, Cacabelos D, Dalfo E et al (2008) Maillard reaction versus other nonenzymatic modifications in neurodegenerative processes. Ann N Y Acad Sci 1126:315319.
  • 86
    Pedersen WA, Fu W, Keller JN, Markesbery WR, Appel S, Smith RG et al (1998) Protein modification by the lipid peroxidation product 4-hydroxynonenal in the spinal cords of amyotrophic lateral sclerosis patients. Ann Neurol 44:819824.
  • 87
    Perez-Gracia E, Torrejon-Escribano B, Ferrer I (2008) Dystrophic neurites of senile plaques in Alzheimer's disease are deficient in cytochrome C oxidase. Acta Neuropathol 116:261268.
  • 88
    Perluigi M, Fai Poon H, Hensley K, Pierce WM, Klein JB, Calabrese V et al (2005) Proteomic analysis of 4-hydroxy-2-nonenal-modified proteins in G93A-SOD1 transgenic mice—a model of familial amyotrophic lateral sclerosis. Free Radic Biol Med 38:960968.
  • 89
    Perluigi M, Poon HF, Maragos W, Pierce WM, Klein JB, Calabrese V et al (2005) Proteomic analysis of protein expression and oxidative modification in r6/2 transgenic mice: a model of Huntington disease. Mol Cell Proteomics 4:18491861.
  • 90
    Petersen RB, Nunomura A, Lee HG, Casadesus G, Perry G, Smith MA, Zhu X (2007) Signal transduction cascades associated with oxidative stress in Alzheimer's disease. J Alzheimers Dis 11:143152.
  • 91
    Petrak J, Ivanek R, Toman O, Cmejla R, Cmejlova J, Vyoral D et al (2008) Déjà vu in proteomics. A hit parade of repeatedly identified differentially expressed proteins. Proteomics 8:17441749.
  • 92
    Pierce A, Mirzaei H, Muller F, De Waal E, Taylor AB, Leonard S et al (2008) GAPDH is conformationally and functionally altered in association with oxidative stress in mouse models of amyotrophic lateral sclerosis. J Mol Biol 382:11951210.
  • 93
    Piert M, Koeppe RA, Giordani B, Berent S, Kuhl DE (1996) Diminished glucose transport and phosphorylation in Alzheimer's disease determined by dynamic FDG-PET. J Nucl Med 37:201208.
  • 94
    Pietrini P, Furey ML, Guazzelli M, Alexander GE (2001) Functional brain studies of the neurometabolic bases of cognitive and behavioral changes in Alzheimer's disease. In: Functional Neurobiology of Aging. PRHof, CVMobbs (eds), pp. 227241. Academic Press: New York.
  • 95
    Poon HF, Frasier M, Shreve N, Calabrese V, Wolozin B, Butterfield DA (2005) Mitochondrial associated metabolic proteins are selectively oxidized in A30P alpha-synuclein transgenic mice—a model of familial Parkinson's disease. Neurobiol Dis 18:492498.
  • 96
    Poon HF, Hensley K, Thongboonkerd V, Merchant ML, Lynn BC, Pierce WM et al (2005) Redox proteomics analysis of oxidatively modified proteins in G93A-SOD1 transgenic mice—a model of familial amyotrophic lateral sclerosis. Free Radic Biol Med 39:453462.
  • 97
    Portero-Otin M, Pamplona R (2006) Is endogenous oxidative protein damage envolved in the aging process? In: Protein Oxidation and Disease. JPietzsch (ed.), pp. 91142. Research Signpost, Kerala, India.
  • 98
    Practico D (2008) Oxidative stress hypothesis in Alzheimer's disease: a reappraisal. Trends Pharmacol Sci 29:609615.
  • 99
    Pryor WA, Squadrito GL (1995) The chemistry of peroxynitrite: a product from the reaction of nitric oxide with superoxide. Am J Physiol 268:L699L722.
  • 100
    Reed T, Perluigi M, Sultana R, Pierce WM, Klein JB, Turner DM et al (2008) Proteomic identification of 4-hydroxy-2-nonenal-modified brain proteins in amnestic mild cognitive impairment: insight into the role of lipid peroxidation in the progression and pathogenesis of Alzheimer's disease. Neurobiol Dis 30:107120.
  • 101
    Reed TT, Pierce WM Jr, Turner DM, Markesbery WR, Butterfield DA (2008) Proteomic identification of nitrated brain proteins in early Alzheimer's disease inferior parietal lobule. J Cell Mol Med DOI: 10.1111/j.1582-4934.2008.00478.
  • 102
    Requena J, Chao CC, Stadtman ER (2001) Glutamic acid and aminodipidic semialdehydes are thye main carbonyl products of metal-catalyzed oxidation of proteins. Proc Natl Acad Sci USA 98:624632.
  • 103
    Santpere G, Ferrer I (2008) Delineation of progressive supranuclear palsy-like pathology. Astrocytes in striatum are primary targets of tau phosphorylation and GFAP oxidation. Brain Pathol 19:177187.
  • 104
    Santpere G, Puig B, Ferrer I (2007) Oxidative damage of 14-3-3 zeta and gamma isoforms in Alzheimer's disease and cerebral amyloid angiopathy. Neuroscience 146:16401651.
  • 105
    Shin SJ, Lee SE, Boo JH, Kim M, Yoon YD, Kim SI, Mook-Jung I (2004) Profiling proteins related to amyloid deposited brain of Tg2576 mice. Proteomics 4:33593368.
  • 106
    Smith MA, Rudnicka-Nawrot M, Richey P, Praprotnik D, Mulvihill P, Miller CA et al (1995) Carbonyl-related post-translational modification of neurofilament protein in the neurofibrillary pathology of Alzheimer's disease. J Neurochem 64:26602666.
  • 107
    Sorolla MA, Reverter-Branchat G, Tamarit J, Ferrer I, Ros J, Cabiscol E (2008) Proteomic and oxidative stress analysis in human brain samples of Huntington disease. Free Radic Biol Med 45:667678.
  • 108
    Stadtman ER (1998) Free radical-mediated oxidation of proteins. In: Free Radicals, Oxidative Stress, and Antioxidants: Pathological and Physiological Significance. NATO ASI Series, Series A: Life Sciences, Vol. 296. TOzben (ed.), pp. 51143. Plenum Press: New York.
  • 109
    Stadtman ER (2002) Importance of individuality in oxidative stress and aging. Free Radic Biol Med 33:597604.
  • 110
    Stadtman ER, Berlett BS (1997) Free radical-mediated modification of proteins. In: Free Radical Toxicity. KBWallace (ed.), pp. 7187. Taylor and Francis: Washington, DC.
  • 111
    Stadtman ER, Levine RL (2006) Chemical modification of proteins by reactive oxygen species. In: Redox Proteomics: From Protein Modifications to Cellular Dysfunction and Diseases. IDalla-Donne, AScaloni, DAButterfield (eds), pp. 323. John Wiley & Sons: Hoboken, NJ.
  • 112
    Starkov AA (2008) The role of mitochondria in reactive oxygen species metabolism and signaling. Ann N Y Acad Sci 1147:3752.
  • 113
    Su B, Wang X, Nunomura A, Moreira PI, Lee HG, Perry G et al (2008) Oxidative stress signalling in Alzheimer's disease. Curr Alzheimer Res 5:525532.
  • 114
    Sultana R, Boyd-Kimball D, Poon HF, Cai J, Pierce WM, Klein JB et al (2006) Oxidative modification and down-regulation of Pin1 in Alzheimer's disease hippocampus: a redox proteomics analysis. Neurobiol Aging 27:918925.
  • 115
    Sultana R, Boyd-Kimball D, Poon HF, Cai J, Pierce WM, Klein JB et al (2006) Redox proteomics identification of oxidized proteins in Alzheimer's disease hippocampus and cerebellum: an approach to understand pathological and biochemical alterations in AD. Neurobiol Aging 27:15641576.
  • 116
    Sultana R, Newman SF, Abdul HM, Cai J, Pierce WM, Klein JB et al (2006) Protective effect of D609 against amyloid-beta1-42-induced oxidative modification of neuronal proteins: redox proteomics study. J Neurosci Res 84:409417.
  • 117
    Sultana R, Poon HF, Cai J, Pierce WM, Merchant M, Klein JB et al (2006) Identification of nitrated proteins in Alzheimer's disease brain using a redox proteomics approach. Neurobiol Dis 22:7687.
  • 118
    Sultana R, Perluigi M, Butterfield DA (2009) Oxidatively modified proteins in Alzheimer's disease (AD), mild cognitive impairment and animal models of AD: role of Abeta in pathogenesis. Acta Neuropathol 118:131150.
  • 119
    Terni B, Boada J, Portero-Otín M, Pamplona R, Ferrer I (2009) Mitochondrial ATP-synthase in the entorhinal cortex is a target of oxidative stress at stages I/II of Alzheimer's disease pathology. Brain Pathol DOI: 10.1111/j.1750-3639.2009.00266.x.
  • 120
    Thannickal VJ, Fanburg BL (2000) Reactive oxygen species in cell signaling. Am J Physiol Lung Cell Mol Physiol 279:L1005L1028.
  • 121
    Thomas DD, Ridnour L, Donzelli S, Espey MG, Mancardi D, Isenberg JS et al (2006) The chemistry of protein modifications elicited by nitric oxide and related nitrogen oxides. In: Redox Proteomics: From Protein Modifications to Cellular Dysfunction and Diseases. IDalle-Done, AScaloni, DAButterfield (eds), pp. 2558. John Wiley & Sons: Hoboken, NJ.
  • 122
    Thorpe SR, Baynes JW (2003) Maillard reaction products in tissue proteins: new products and new perspectives. Amino Acids 25:275281.
  • 123
    Wataya T, Nunomura A, Smith MA, Siedlak SL, Harris PLR, Shimohama S et al (2002) High molecular weight neurofilament proteins are physiological substrates of aduction by the lipid peroxidation product hydroxynonenal. J Biol Chem 277:46444648.
  • 124
    Wolff SP, Jiang ZY, Hunt JV (1991) Protein glycation and oxidative stress in diabetes mellitus and ageing. Free Radic Biol Med 10:339352.
  • 125
    Wong-Riley M, Antuono P, Ho KV, Egan R, Hevner R, Liebl WW et al (1997) Cytochrome oxiudase in Alzheimer's disease: biochemical, histochemical, and immunohistochemical analysis of the visual and other systems. Vision Res 37:35933608.
  • 126
    Yao D, Gu Z, Nakamura T, Shi ZQ, Ma Y, Gaston B et al (2004) Nitrosative stress linked to sporadic Parkinson's disease: S-nitrosylation of parkin regulates its E3 ubiquitin ligase activity. Proc Natl Acad Sci USA 101:1081010814.
  • 127
    Zhou C, Huang Y, Przedborski S (2008) Oxidative stress in Parkinson's disease: a mechanism of pathogenic and therapeutic significance. Ann N Y Acad Sci 1147:93104.
  • 128
    Zhu X, Raina AK, Lee HG, Casadesus G, Smith MA, Perry G (2004) Oxidative stress signalling in Alzheimer's disease. Brain Res 1000:3239.
  • 129
    Zhu X, Lee HG, Casadesus G, Avila J, Drew K, Perry G, Smith MA (2005) Oxidative imbalance in Alzheimer's disease. Mol Neurobiol 31:205217.
  • 130
    Zhu X, Su B, Wang X, Smith MA, Perry G (2007) Causes of oxidative stress in Alzheimer's disease. Cell Mol Life Sci 64:22022210.

Supporting Information

  1. Top of page
  2. Abstract
  3. INTRODUCTION
  4. IDENTIFICATION OF OXIDIZED AND NITRATED PROTEINS
  5. ALZHEIMER DISEASE
  6. TAUOPATHIES
  7. PARKINSON DISEASE AND RELATED α-SYNUCLEINOPATHIES
  8. HUNTINGTON DISEASE
  9. AMYOTROPHIC LATERAL SCLEROSIS
  10. COMMON AND SPECIFIC PROTEIN TARGETS IN DIFFERENT PATHOLOGIES
  11. OXIDATIVE CHANGES IN POST-MORTEM BRAIN ARE PRINCIPALLY PRIMARY
  12. PROTEIN OXIDATION IS AN EARLY EVENT IN NEURODEGENERATIVE DISEASES
  13. OXIDATIVE DAMAGE AND LOSS OF FUNCTION
  14. CLINICAL IMPLICATIONS
  15. CELLULAR LOCALIZATION OF OXIDATIVELY DAMAGED PROTEINS
  16. PITFALLS AND LIMITATIONS
  17. REFINING METHODS TO IMPROVE REDOX PROTEOMICS
  18. CONCLUDING COMMENTS
  19. ACKNOWLEDGMENTS
  20. REFERENCES
  21. Supporting Information

Table S1. Alzheimer disease.

Table S2. Tauopathies.

Table S3. Parkinson disease and related α-synucleinopathies.

Please note: Wiley-Blackwell are not responsible for the content or functionality of any supporting materials supplied by the authors. Any queries (other than missing material) should be directed to the corresponding author for the article.

FilenameFormatSizeDescription
BPA_326_sm_table_1.doc83KSupporting info item
BPA_326_sm_table_2.doc33KSupporting info item
BPA_326_sm_table_3.doc45KSupporting info item

Please note: Wiley Blackwell is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.