Kinetic and thermodynamic properties of the folding and assembly of formate dehydrogenase.
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 2x10(-3)s(-1) (by deduction k(-1) is about 10(-4)s(-1)) and assembles into the active dimeric state with a bimolecular rate constant (k(2)) of about 2x10(4)M(-1)s(-1). The rate of dissociation of the dimeric state in physiological conditions is extremely slow (k(-2) approximately 3x10(-7)s(-1)).
Ordu
Emel B
EB
Istanbul Technical University, Faculty of Science and Letters, Department of Molecular Biology and Genetics, Istanbul, Turkey.
Cameron
Gus
G
Clarke
Anthony R
AR
Karagüler
Nevin Gül
NG
D002175:Candida / Q000201:enzymology; D005560:Formate Dehydrogenases / Q000737:chemistry* / Q000378:metabolism; D005656:Fungal Proteins / Q000737:chemistry* / Q000378:metabolism; D006358:Hot Temperature; D007700:Kinetics; D011489:Protein Denaturation; D017510:Protein Folding; D055503:Protein Multimerization; D020836:Protein Structure, Quaternary; D013816:Thermodynamics
D005656:Fungal Proteins; D005560:Formate Dehydrogenases