FEBS Letters
Explore this journal >
Short communication

Kinetic and thermodynamic properties of the folding and assembly of formate dehydrogenase

Authors

  • Emel B. Ordu,

    1. Istanbul Technical University, Faculty of Science and Letters, Department of Molecular Biology and Genetics, Istanbul, Turkey
    2. Istanbul Technical University, Molecular Biology-Biotechnology and Genetics Research Centre, 34469 Istanbul, Turkey
    3. Yildiz Technical University, Faculty of Science and Letters, Department of Biology, Davutpasa, 34220 Esenler, Istanbul, Turkey
    Search for more papers by this author

  • Gus Cameron,

    1. University of Bristol, Department of Biochemistry, Bristol BS8 1TD, Avon, England
    Search for more papers by this author

  • Anthony R. Clarke,

    1. University of Bristol, Department of Biochemistry, Bristol BS8 1TD, Avon, England
    Search for more papers by this author

  • Nevin Gül Karagüler

    Corresponding author
    1. Istanbul Technical University, Faculty of Science and Letters, Department of Molecular Biology and Genetics, Istanbul, Turkey
    2. Istanbul Technical University, Molecular Biology-Biotechnology and Genetics Research Centre, 34469 Istanbul, Turkey

    • Corresponding author. Address: Istanbul Technical University, Faculty of Science and Letters, Department of Molecular Biology and Genetics, 34469 Istanbul, Turkey. Fax: +90 212 2856386.

    Search for more papers by this author

Abstract

The folding mechanism and stability of dimeric formate dehydrogenase from Candida methylica was analysed by exposure to denaturing agents and to heat. Equilibrium denaturation data yielded a dissociation constant of about 10−13 M for assembly of the protein from unfolded chains and the kinetics of refolding and unfolding revealed that the overall process comprises two steps. In the first step a marginally stable folded monomeric state is formed at a rate (k 1) of about 2 × 10−3 s−1 (by deduction k −1 is about10−4 s−1) and assembles into the active dimeric state with a bimolecular rate constant (k 2) of about 2 × 104 M−1 s−1. The rate of dissociation of the dimeric state in physiological conditions is extremely slow (k −2 ∼ 3 × 10−7 s−1).

Continue reading full article

Ancillary

Article Information

DOI

10.1016/j.febslet.2009.07.048

Format Available

Full text: HTML | PDF

FEBS Letters 583 (2009) 1873-3468 © 2015 Federation of European Biochemical Societies

Request Permissions

Keywords

  • cmFDH;
  • Candida metylica formate dehydrogenase;
  • NAD;
  • nicotinamide adenine dinucleotide;
  • yTIM;
  • yeast triosephosphate isomerase;
  • apo-SOD;
  • apo-superoxide dismutase;
  • N-PGK;
  • N-domain of phosphoglycerate kinase;
  • Assembly mechanism;
  • Folding mechanism;
  • Formate dehydrogenase;
  • Candida methylica

Publication History

  • Issue online:
  • Version of record online:
  • Manuscript Accepted:
  • Manuscript Revised:
  • Manuscript Received:

Supporting Information

FilenameDescription
feb2s0014579309005985-sup-fx1.jpgapplication/jpg, 18K

Supplementary Fig. 1. Thermodynamic analysis of the unfolding barrier. Samples of FDH (50 μM) were incubated at the temperatures shown and aliquots removed for assay at a series of times. ○ The resultant data (residual activity v time) were well described by single-exponential decays and the rate constants were extracted. ○ The plot show rate constant as a function of temperature fitted to the equation:

display math

where

display math

○ as described in Section 2.

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.

Related content

Articles related to the one you are viewing

Citing Literature

  • Number of times cited: 7
  1. 1Dilek Alagöz, Ayhan Çelik, Deniz Yildirim, S. Seyhan Tükel, Barış Binay, Covalent immobilization of Candida methylica formate dehydrogenase on short spacer arm aldehyde group containing supports, Journal of Molecular Catalysis B: Enzymatic, 2016, 130, 40CrossRef
  2. 2Guzman Torrelo, Ulf Hanefeld, Frank Hollmann, Biocatalysis, Catalysis Letters, 2015, 145, 1, 309CrossRef
  3. 3Guzman Torrelo, Ulf Hanefeld, Frank Hollmann, Kirk-Othmer Encyclopedia of Chemical Technology, 2015, 1Wiley Online Library
  4. 4Emel B. Ordu, Richard B. Sessions, Anthony R. Clarke, Nevin Gül Karagüler, Effect of surface electrostatic interactions on the stability and folding of formate dehydrogenase from Candida methylica, Journal of Molecular Catalysis B: Enzymatic, 2013, 95, 23CrossRef
  5. 5Emel B. Ordu, Emrah Yelboğa, Francesco Secundo, Richard B. Sessions, Nevin Gül Karagüler, The effect of methionine to cysteine substitution on the stability of formate dehydrogenase from Candida methylica, Journal of Molecular Catalysis B: Enzymatic, 2012, 82, 109CrossRef
  6. 6Frank Hollmann, Isabel W. C. E. Arends, Katja Buehler, Biocatalytic Redox Reactions for Organic Synthesis: Nonconventional Regeneration Methods, ChemCatChem, 2010, 2, 7, 762Wiley Online Library
  7. 7Makoto Yoshimoto, Ryo Yamasaki, Manami Nakao, Takayuki Yamashita, Stabilization of formate dehydrogenase from Candida boidinii through liposome-assisted complexation with cofactors, Enzyme and Microbial Technology, 2010, 46, 7, 588CrossRef