Zinc binding modulates the entire folding free energy surface of human Cu,Zn superoxide dismutase.
Biochemistry & Molecular Pharmacology
Department of Biochemistry and Molecular Pharmacology
Biochemistry, Biophysics, and Structural Biology | Life Sciences | Medicine and Health Sciences
Over 100 amino acid replacements in human Cu,Zn superoxide dismutase (SOD) are known to cause amyotrophic lateral sclerosis, a gain-of-function neurodegenerative disease that destroys motor neurons. Supposing that aggregates of partially folded states are primarily responsible for toxicity, we determined the role of the structurally important zinc ion in defining the folding free energy surface of dimeric SOD by comparing the thermodynamic and kinetic folding properties of the zinc-free and zinc-bound forms of the protein. The presence of zinc was found to decrease the free energies of a peptide model of the unfolded monomer, a stable variant of the folded monomeric intermediate, and the folded dimeric species. The unfolded state binds zinc weakly with a micromolar dissociation constant, and the folded monomeric intermediate and the native dimeric form both bind zinc tightly, with subnanomolar dissociation constants. Coupled with the strong driving force for the subunit association reaction, the shift in the populations toward more well-folded states in the presence of zinc decreases the steady-state populations of higher-energy states in SOD under expected in vivo zinc concentrations (approximately nanomolar). The significant decrease in the population of partially folded states is expected to diminish their potential for aggregation and account for the known protective effect of zinc. The approximately 100-fold increase in the rate of folding of SOD in the presence of micromolar concentrations of zinc demonstrates a significant role for a preorganized zinc-binding loop in the transition-state ensemble for the rate-limiting monomer folding reaction in this beta-barrel protein.
DOI of Published Version
J Mol Biol. 2008 Dec 12;384(2):540-55. Epub 2008 Sep 26. Link to article on publisher's website
Journal of molecular biology
Kayatekin, Can; Zitzewitz, Jill A.; and Matthews, C. Robert, "Zinc binding modulates the entire folding free energy surface of human Cu,Zn superoxide dismutase." (2008). GSBS Student Publications. 1592.