ChemBioChem

Cover image for Vol. 10 Issue 10

July 6, 2009

Volume 10, Issue 10

Pages 1577–1739

  1. Cover Picture

    1. Top of page
    2. Cover Picture
    3. Graphical Abstract
    4. News
    5. Minireview
    6. Highlights
    7. Communications
    8. Full Papers
    9. Book Review
    10. Preview
    1. Cover Picture: Analysis of Protein–Protein Interactions by Using Droplet-Based Microfluidics (ChemBioChem 10/2009) (page 1577)

      Monpichar Srisa-Art, Dong-Ku Kang, Jongin Hong, Hyun Park, Robin J. Leatherbarrow, Joshua B. Edel, Soo-Ik Chang and Andrew J. deMello

      Article first published online: 29 JUN 2009 | DOI: 10.1002/cbic.200990037

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      The cover picture shows a segmented-flow microfluidic system that was used to analyse protein–protein interactions in picolitre droplets. The 400 pL aqueous droplets were formed, encapsulated by a carrier fluid, and then transported through a 50 μm-wide microchannel at a constant velocity. Each droplet contained fluorescently labelled antigens and antibodies, and FRET was used to report protein–protein interactions. Angiogenin (ANG), a small polypeptide implicated in angiogenesis and tumour growth, was selected as a model protein. Specifically, an anti-ANG antibody (anti-ANG Ab) and an ANG antigen were labelled with fluorophores to act as donor and acceptor in the FRET measurements. KD values for ANG and anti-ANG Ab from these experiments (KD=6.4±1.6 nM) agree closely with data from bulk fluorescence polarisation measurements KD=9.0±1.5 nM). We expect that this novel experimental platform will have significant application in high-throughput protein expression profiling and drug discovery. For further details, see the article by S.-I. Chang, A. J. deMello et al. on p. 1605 ff.

  2. Graphical Abstract

    1. Top of page
    2. Cover Picture
    3. Graphical Abstract
    4. News
    5. Minireview
    6. Highlights
    7. Communications
    8. Full Papers
    9. Book Review
    10. Preview
    1. Graphical Abstract: ChemBioChem 10/2009 (pages 1579–1586)

      Article first published online: 29 JUN 2009 | DOI: 10.1002/cbic.200990038

  3. News

    1. Top of page
    2. Cover Picture
    3. Graphical Abstract
    4. News
    5. Minireview
    6. Highlights
    7. Communications
    8. Full Papers
    9. Book Review
    10. Preview
  4. Minireview

    1. Top of page
    2. Cover Picture
    3. Graphical Abstract
    4. News
    5. Minireview
    6. Highlights
    7. Communications
    8. Full Papers
    9. Book Review
    10. Preview
    1. The Polarised Life of the Endocannabinoid System in CNS Development (pages 1591–1598)

      Sharon Anavi-Goffer and Jan Mulder

      Article first published online: 16 JUN 2009 | DOI: 10.1002/cbic.200800827

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      The spatiotemporal expression of cannabinoid receptors and endocannabinoid-metabolising enzymes during brain development guides major developmental processes including neurogenesis, cell differentiation, cell migration, neuronal specification and synaptogenesis.

  5. Highlights

    1. Top of page
    2. Cover Picture
    3. Graphical Abstract
    4. News
    5. Minireview
    6. Highlights
    7. Communications
    8. Full Papers
    9. Book Review
    10. Preview
    1. Expanding RNA Silencing Approaches by U1 Adaptors (pages 1599–1601)

      Arnold Grünweller and Roland K. Hartmann

      Article first published online: 16 JUN 2009 | DOI: 10.1002/cbic.200900271

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      Silent partner: In a recent publication, Gunderson and co-workers described an approach to recruit the abundant U1 snRNP, a splicing subparticle, to the last exon of a pre-mRNA, in a sequence-specific manner. This mediates interaction of the U1-70K protein subunit with poly(A) polymerase, thus blocking polyadenylation and inducing pre-mRNA degradation. This novel promising strategy expands the repertoire of gene-silencing concepts.

    2. Hacking the Genetic Code of Mammalian Cells (pages 1602–1604)

      Dirk Schwarzer

      Article first published online: 16 JUN 2009 | DOI: 10.1002/cbic.200900302

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      A genetic shuttle: The highlighted article, which was recently published by Schultz, Geierstanger and co-workers, describes a straightforward scheme for enlarging the genetic code of mammalian cells. An orthogonal tRNA/aminoacyl-tRNA synthetase pair specific for a new amino acid can be evolved in E. coli and subsequently transferred into mammalian cells. The feasibility of this approach was demonstrated by adding a photocaged lysine derivative to the genetic repertoire of a human cell line.

  6. Communications

    1. Top of page
    2. Cover Picture
    3. Graphical Abstract
    4. News
    5. Minireview
    6. Highlights
    7. Communications
    8. Full Papers
    9. Book Review
    10. Preview
    1. Analysis of Protein–Protein Interactions by Using Droplet-Based Microfluidics (pages 1605–1611)

      Monpichar Srisa-Art, Dong-Ku Kang, Jongin Hong, Hyun Park, Robin J. Leatherbarrow, Joshua B. Edel, Soo-Ik Chang and Andrew J. deMello

      Article first published online: 3 JUN 2009 | DOI: 10.1002/cbic.200800841

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      Every little drop: The KD values of angiogenin (ANG) interactions as shown by FRET analysis of thousands of pL-sized droplets agree with data from bulk-fluorescence polarization measurements. Importantly, the use of fluorophores does not affect the activity of ANG or the binding of anti-ANG antibodies to ANG. Such an experimental platform could be applied to the high-throughput analysis of protein–protein interactions.

    2. Photochemical Regulation of Restriction Endonuclease Activity (pages 1612–1616)

      Douglas D. Young, Jeane M. Govan, Mark O. Lively and Alexander Deiters

      Article first published online: 16 JUN 2009 | DOI: 10.1002/cbic.200900090

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      Removal by the light: The photochemical regulation of restriction endonucleases, which are important enzymes in molecular biology, has been investigated. Photolabile protecting groups have been installed on DNA substrates and have been demonstrated to inhibit restriction endonuclease activity until removed by UV light irradiation. Interestingly, these groups do not appear to dramatically affect initial binding of the enzyme to the DNA substrate, but rather prevent recognition of the specific cleavage site.

    3. Synthesis and Investigation of Tryptophan–Amphotericin B Conjugates (pages 1617–1620)

      Andreas Zumbuehl, Pasquale Stano, Marc Sohrmann, Rolf Dietiker, Mathias Peter and Erick M. Carreira

      Article first published online: 16 JUN 2009 | DOI: 10.1002/cbic.200900096

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      Anchors aweigh! The synthesis of tryptophan–amphotericin B conjugates (see figure) is described. The membrane-anchoring effect of tryptophane was thus combined with the pore-formation effect of amphotericin B leading to high channel activity in sterol-free liposomes.

    4. Eosin Y-Sensitized Artificial Photosynthesis by Highly Efficient Visible-Light-Driven Regeneration of Nicotinamide Cofactor (pages 1621–1624)

      Sahng Ha Lee, Dong Heon Nam, Jae Hong Kim, Jin-Ook Baeg and Chan Beum Park

      Article first published online: 23 JUN 2009 | DOI: 10.1002/cbic.200900156

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      Dye-sensitized photosynthesis: Eosin Y (EY), a dye photosensitizer, works efficiently as a molecular photoelectrode by catalyzing the visible-light-driven electron-transfer reaction for efficient regeneration of NADH through a photosensitizer–electron relay dyad. Injection of the photosensitized electron resulted in highly accelerated regeneration of NADH, which can be used by glutamate dehydrogenase for the photosynthesis of L-glutamate.

    5. Monoclonal Antibodies with Orthogonal Azaspiracid Epitopes (pages 1625–1629)

      Michael O. Frederick , Sandra De Lamo Marin, Kim D. Janda, K. C. Nicolaou  and Tobin J. Dickerson

      Article first published online: 2 JUN 2009 | DOI: 10.1002/cbic.200900201

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      Azaspiracid antibodies: Immunization of azaspiracid immunoconjugates has elicited monoclonal antibodies with distinct epitopes on the marine toxin; this will open the way toward azaspiracid diagnostics and the detection of contaminated shellfish before they can enter the food supply.

    6. Steric Constraints Dependent on Nucleobase Pair Orientation Vary in Different DNA Polymerase Active Sites (pages 1630–1633)

      Frank Streckenbach, Gopinath Rangam, Heiko M. Möller and Andreas Marx

      Article first published online: 18 JUN 2009 | DOI: 10.1002/cbic.200900123

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      Finding the right fit: Herein, we report on the development of novel steric probes and present initial insights into their interplay with DNA polymerases. Our findings provide experimental evidence for varied enzyme–substrate interactions that might account for the varied selectivity previously observed.

    7. Combinatorial Strategies by Marine Cyanobacteria: Symplostatin 4, an Antimitotic Natural Dolastatin 10/15 Hybrid that Synergizes with the Coproduced HDAC Inhibitor Largazole (pages 1634–1639)

      Kanchan Taori, Yanxia Liu, Valerie J. Paul and Hendrik Luesch

      Article first published online: 9 JUN 2009 | DOI: 10.1002/cbic.200900192

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      Combinatorial biosynthesis meets combinatorial pharmacology, cyanobacterial style: A new antimitotic natural product with features of both dolastatins 10 and 15 was isolated from the same Floridian Symploca sp. sample that produced the histone deacetylase inhibitor largazole. Both agents in combination are more effective in inhibiting cancer cell proliferation than either agent alone.

    8. Time-Resolved Tracking of a Minimum Gene Expression System Reconstituted in Giant Liposomes (pages 1640–1643)

      Hirohide Saito, Yusho Kato, Maël Le Berre, Ayako Yamada, Tan Inoue, Kenichi Yosikawa and Damien Baigl

      Article first published online: 16 JUN 2009 | DOI: 10.1002/cbic.200900205

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      Individual expression: We describe a method that allows the observation of real-time gene expression in a large number of individual giant liposomes encapsulating identical genetic material. We followed the gene expression profiles from DNA and mRNA templates coding for different proteins. Although the average profiles of individual liposomes were similar to those measured in bulk solution, strong variability between individual liposomes was observed at both transcription and translation.

    9. Bipartite Tetracysteine Display Requires Site Flexibility for ReAsH Coordination (pages 1644–1647)

      Jessica L. Goodman, Daniel B. Fried and Alanna Schepartz

      Article first published online: 16 JUN 2009 | DOI: 10.1002/cbic.200900207

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      Flexibility required: We designed intramolecular bipartite tetracysteine sites in loops of p53 and the β-sheets of EmGFP. We found that ReAsH binding preferentially favors tetracysteine sites with flexible geometries such as loops; flexibility was assessed by comparing Cα B-factor values. This information is important for directing successful bipartite tetracysteine site designs.

    10. Spatio-Temporal Control of Cell Coculture Interactions on Surfaces (pages 1648–1653)

      Eun-Ju Lee, Eugene W. L. Chan and Muhammad N. Yousaf

      Article first published online: 23 JUN 2009 | DOI: 10.1002/cbic.200900277

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      Coculture control: We report a combined photochemical and electroactive self-assembled monolayer (SAM)-based substrate strategy to generate a coculture platform with spatial and temporal control of cell–cell interactions. These dynamic substrates can present a variety of ligands on the surface for biospecific interactions between the ligands and cell surface receptors. Photopatterning enables the ligands to be immobilized in patterns and even gradients.

    11. A Highly Active Single-Mutation Variant of P450BM3 (CYP102A1) (pages 1654–1656)

      Christopher J. C. Whitehouse, Stephen G. Bell, Wen Yang, Jake A. Yorke, Christopher F. Blanford, Anthony J. F. Strong, Edward J. Morse, Mark Bartlam, Zihe Rao and Luet-Lok Wong

      Article first published online: 2 JUN 2009 | DOI: 10.1002/cbic.200900279

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      The power of proline: Bold amino acid substitutions in sensitive protein regions are frequently unproductive, while more subtle mutations can be sufficient to bring about dramatic changes. But introducing proline at the residue next to the sulfur ligand in P450BM3 (CYP102A1) has the unexpected and desirable effect of enhancing the activity of this fatty acid hydroxylase with a broad range of non-natural substrates, as illustrated by the figure.

  7. Full Papers

    1. Top of page
    2. Cover Picture
    3. Graphical Abstract
    4. News
    5. Minireview
    6. Highlights
    7. Communications
    8. Full Papers
    9. Book Review
    10. Preview
    1. A Combined Solid-State NMR and MD Characterization of the Stability and Dynamics of the HET-s(218-289) Prion in its Amyloid Conformation (pages 1657–1665)

      Adam Lange, Zrinka Gattin, Hélène Van Melckebeke, Christian Wasmer, Alice Soragni, Wilfred F. van Gunsteren and Beat H. Meier

      Article first published online: 5 JUN 2009 | DOI: 10.1002/cbic.200900019

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      Dynamic and rigid: The prion HET-s(218–289) consists, in its amyloid form as shown here, of highly ordered and rigid parts and a very dynamic loop, which could be of great importance for fibril formation. Indeed, MD simulations explain the experimental NMR results and describe the dynamics of the salt-bridge network that stabilizes the amyloid fibril, a feature not easily accessible by experiment.

    2. Aphrodisiac Pheromones from the Wings of the Small Cabbage White and Large Cabbage White Butterflies, Pieris rapae and Pieris brassicae (pages 1666–1677)

      Selma Yildizhan, Joop van Loon, Anna Sramkova, Manfred Ayasse, Cristian Arsene, Cindy ten Broeke and Stefan Schulz

      Article first published online: 16 JUN 2009 | DOI: 10.1002/cbic.200900183

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      Pheromonally yours: The aphrodisiac pheromones of male Cabbage White butterflies were identified. The small butterfly uses the smaller molecule ferrulactone (1) to enhance its mating success, while the large butterfly uses the larger compound brassicalactone (2), which is a new natural product. Both compounds are active in combination with hexahydrofarnesylacetone and phytol.

    3. Development of a Method for the High-Throughput Quantification of Cellular Proteins (pages 1678–1688)

      Paolo Paganetti, Andreas Weiss , Monique Trapp, Ina Hammerl, Dorothée Bleckmann, Ruth A. Bodner, Shanie Coven-Easter, David E. Housman and Christian N. Parker

      Article first published online: 2 JUN 2009 | DOI: 10.1002/cbic.200900131

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      Hunting for huntingtin: We describe a screening assay based on the inducible expression of the mutant huntingtin protein in cells and on its highly sensitive homogenous determination. Rapid, reproducible, and robust protein determination was achieved through the use of two donor–acceptor-labeled antibodies and time-resolved FRET. The assay was developed and validated for ultra-throughput screening of low-molecular-weight compounds modulating the expression of the mutant protein.

    4. Small-Molecule Targeting of the Mitochondrial Compartment with an Endogenously Cleaved Reversible Tag (pages 1689–1696)

      Jens Ripcke, Kim Zarse, Michael Ristow and Marc Birringer

      Article first published online: 2 JUN 2009 | DOI: 10.1002/cbic.200900159

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      Reversible mitochondrial shuttle: A novel concept in mitochondrial pharmacology allows the transport of bioactive compounds into the mitochondrial compartment and their subsequent release. A lipoic acid derivative containing a cleavable (“reversible”) triphenylphosphonium tag is endogenously cleaved by the mitochondrial aldehyde dehydrogenase (ALDH-2) after mitochondrial accumulation.

    5. Epoxide-Hydrolase-Initiated Hydrolysis/Rearrangement Cascade of a Methylene-Interrupted Bis-Epoxide Yields Chiral THF Moieties without Involvement of a “Cyclase” (pages 1697–1704)

      Barbara T. Ueberbacher, Gustav Oberdorfer, Karl Gruber and Kurt Faber

      Article first published online: 3 JUN 2009 | DOI: 10.1002/cbic.200900176

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      Cyclase-free at last: A methylene-interrupted meso-bis-epoxide was stereoselectively converted into dihydroxy-tetrahydrofuran derivatives with excellent de and ee values through an enzyme-triggered nucleophilic hydrolysis/cyclisation cascade. Molecular modelling showed that the point of enzyme attack was determined by the stereospecificity of the epoxide hydrolase, whereas the stereochemical course of the cyclisation step was solely governed by Baldwin's rules and did not invoke the involvement of a “cyclase”.

    6. Improvement of Yarrowia lipolytica Lipase Enantioselectivity by Using Mutagenesis Targeted to the Substrate Binding Site (pages 1705–1713)

      F. Bordes, E. Cambon, V. Dossat-Létisse, I. André, C. Croux, J. M. Nicaud and A. Marty

      Article first published online: 5 JUN 2009 | DOI: 10.1002/cbic.200900215

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      Enhanced enantioselectivity: The resolution of 2-bromo-arylacetic acid esters by Lip2p lipase from Yarrowia lipolytica was improved through mutagenesis of the substrate binding site. Position 232 was identified as crucial for the discrimination. Saturation of this position led to the identification of variant V232S, which has a tremendously increased activity and E value as compared to the parental enzyme.

    7. Malonyl carba(dethia)- and Malonyl oxa(dethia)-coenzyme A as Tools for Trapping Polyketide Intermediates (pages 1714–1723)

      Manuela Tosin, Dieter Spiteller and Jonathan B. Spencer

      Article first published online: 8 JUN 2009 | DOI: 10.1002/cbic.200900093

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      Caught in a trap. In this study trapped polyketide species (see figure) were off-loaded from a type III PKS by novel nonhydrolyzable malonyl coenzyme A analogues in which a methylene group or an oxygen atom replaces the sulfur atom of malonyl-CoA. This strategy allows the straightforward characterisation of intermediates of polyketide biosynthesis by LC-HR-ESI-MS/MS and provides valuable insights on the mechanism and timing of polyketide formation.

    8. Synthesis and Evaluation of New Thiodigalactoside-Based Chemical Probes to Label Galectin-3 (pages 1724–1733)

      Monique van Scherpenzeel, Ed E. Moret, Lluis Ballell, Rob M. J. Liskamp, Ulf J. Nilsson, Hakon Leffler and Roland J. Pieters

      Article first published online: 2 JUN 2009 | DOI: 10.1002/cbic.200900198

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      Light up galectin: Photoprobes based on thiodigalactoside were prepared for galectin-3, a lectin linked to cancer. The probes contained either benzophenone or acetophenone moieties as the photolabel for covalent attachment to the protein. One particular probe labeled galectin-3 selectively, even in the presence of cell lysate.

  8. Book Review

    1. Top of page
    2. Cover Picture
    3. Graphical Abstract
    4. News
    5. Minireview
    6. Highlights
    7. Communications
    8. Full Papers
    9. Book Review
    10. Preview
    1. Modern Biocatalysis: Stereoselective and Environmentally Friendly Reactions. Edited by Wolf-Dieter Fessner and Thorleif Anthonsen. (page 1734)

      Nicholas Turner

      Article first published online: 29 JUN 2009 | DOI: 10.1002/cbic.200900340

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      Wiley-VCH, Weinheim 2009, XXIII+375 pp., € 129.00.—ISBN 978-3-527-32071-4

  9. Preview

    1. Top of page
    2. Cover Picture
    3. Graphical Abstract
    4. News
    5. Minireview
    6. Highlights
    7. Communications
    8. Full Papers
    9. Book Review
    10. Preview
    1. Preview: ChemBioChem 11/2009 (page 1739)

      Article first published online: 29 JUN 2009 | DOI: 10.1002/cbic.200990040

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