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

Ordu E. , Cameron G., Clarke A. R. , Karaguler N. G.

FEBS LETTERS, vol.583, no.17, pp.2887-2892, 2009 (Journal Indexed in SCI) identifier identifier identifier

  • Publication Type: Article / Article
  • Volume: 583 Issue: 17
  • Publication Date: 2009
  • Doi Number: 10.1016/j.febslet.2009.07.048
  • Title of Journal : FEBS LETTERS
  • Page Numbers: pp.2887-2892

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 x 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 x 10(4) M (1) s (1). The rate of dissociation of the dimeric state in physiological conditions is extremely slow (k (2) similar to 3 x 10 (7) s (1)). (C) 2009 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.