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

Emel B. Ordu; Gus Cameron; Anthony R. Clarke; Nevin Gül Karagüler

doi:10.1016/j.febslet.2009.07.048

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

Emel B. Ordu, Gus Cameron, Anthony R. Clarke, Nevin Gül Karagüler^*

^*Corresponding author for this work

Department of Molecular Biology and Genetics

Research output: Contribution to journal › Article › peer-review

10 Citations (Scopus)

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 (k₁) 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 (k₂) 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).

Original language	English
Pages (from-to)	2887-2892
Number of pages	6
Journal	FEBS Letters
Volume	583
Issue number	17
DOIs	https://doi.org/10.1016/j.febslet.2009.07.048
Publication status	Published - 3 Sept 2009

Keywords

Assembly mechanism
Candida methylica
Folding mechanism
Formate dehydrogenase

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10.1016/j.febslet.2009.07.048

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title = "Kinetic and thermodynamic properties of the folding and assembly of formate dehydrogenase",

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 (k1) 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 (k2) 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).",

keywords = "Assembly mechanism, Candida methylica, Folding mechanism, Formate dehydrogenase",

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AU - Ordu, Emel B.

AU - Cameron, Gus

AU - Clarke, Anthony R.

AU - Karagüler, Nevin Gül

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AB - 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 (k1) 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 (k2) 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).

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JO - FEBS Letters

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ER -

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

Abstract

Keywords

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