Unfolding and Folding Kinetics of Amyotrophic Lateral Sclerosis-Associated Mutant Cu,Zn Superoxide Dismutases
| Autor: | Elizabeth M. Meiering, Jessica A.O. Rumfeldt, James R. Lepock |
|---|---|
| Rok vydání: | 2009 |
| Předmět: |
Protein Denaturation
Protein Folding Dimer Kinetics Mutant Metal Superoxide dismutase chemistry.chemical_compound Structural Biology Enzyme Stability medicine Humans Amyotrophic lateral sclerosis Molecular Biology Guanidine biology Superoxide Dismutase Amyotrophic Lateral Sclerosis Hydrogen-Ion Concentration medicine.disease Zinc Monomer chemistry Biochemistry visual_art visual_art.visual_art_medium biology.protein Mutant Proteins Protein folding Protein Multimerization Apoproteins Copper |
| Zdroj: | Journal of Molecular Biology. 385:278-298 |
| ISSN: | 0022-2836 |
| DOI: | 10.1016/j.jmb.2008.10.003 |
| Popis: | More than 110 mutations in dimeric, Cu,Zn superoxide dismutase (SOD) have been linked to the fatal neurodegenerative disease, amyotrophic lateral sclerosis (ALS). In both human patients and mouse model studies, protein misfolding has been implicated in disease pathogenesis. A central step in understanding the misfolding/aggregation mechanism of this protein is the elucidation of the folding pathway of SOD. Here we report a systematic analyses of unfolding and folding kinetics using single- and double-jump experiments as well as measurements as a function of guanidium chloride, protein, and metal concentration for fully metallated (holo) pseudo wild-type and ALS-associated mutant (E100G, G93R, G93A, and metal binding mutants G85R and H46R) SODs. The kinetic mechanism for holo SODs involves native dimer, monomer intermediate, and unfolded monomer, with variable metal dissociation from the monomeric states depending on solution conditions. The effects of the ALS mutations on the kinetics of the holoproteins in guanidium chloride are markedly different from those observed previously for acid-induced unfolding and for the unmetallated (apo) forms of the proteins. The mutations decrease the stability of holo SOD mainly by increasing unfolding rates, which is particularly pronounced for the metal-binding mutants, and have relatively smaller effects on the observed folding kinetics. Mutations also seem to favour increased formation of a Zn-free monomer intermediate, which has been implicated in the formation of toxic aggregates. The results reveal the kinetic basis for the extremely high stability of wild-type holo SOD and the possible consequences of kinetic changes for disease. |
| Databáze: | OpenAIRE |
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