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Kinetic and thermodynamic properties of the folding and assembly of formate dehydrogenase
Author(s) -
Ordu Emel B.,
Cameron Gus,
Clarke Anthony R.,
Karagüler Nevin Gül
Publication year - 2009
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2009.07.048
Subject(s) - chemistry , kinetics , reaction rate constant , formate , dissociation (chemistry) , protein folding , formate dehydrogenase , crystallography , monomer , dissociation constant , biochemistry , organic chemistry , polymer , catalysis , quantum mechanics , physics , receptor
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 × 10 4 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 ).