title: Early steps in oxidation-induced SOD1 misfolding: implications for non-amyloid protein aggregation in familial ALS.
abstract: Among the diseases of protein misfolding, amyotrophic lateral sclerosis (ALS) is unusual in that the proteinaceous neuronal inclusions that are the hallmark of the disease have neither the classic fibrillar appearance of amyloid by transmission electron microscopy nor the affinity for the dye Congo red that is a defining feature of amyloid. Mutations in the Cu, Zn superoxide dismutase (SOD1) cause the largest subset of inherited ALS cases. The mechanism by which this highly stable enzyme misfolds to form non-amyloid aggregates is currently poorly understood, as are the stresses that initiate misfolding. The oxidative damage hypothesis proposes that SOD1's normal free radical scavenger role puts it at risk of oxidative damage and that it is this damage that triggers the misfolding primed by mutation. Here, we present evidence that hydrogen peroxide treatment, which generates free radical species at the SOD1 active site, causes oxidative damage to active-site histidine residues, leading to major structural changes and non-amyloid aggregation similar to that seen in ALS. Time-resolved measurements of release of bound metal ligands, exposure of hydrophobic surface area, and alterations in the SOD1 proton NMR spectrum have allowed us to model the early structural changes occurring as SOD1 misfolds, prior to aggregation. ALS-causing SOD1 mutations apparently alter this pathway by increasing exposure of buried epitopes in misfolded species populated at endpoint. We have identified a well-populated early misfolding intermediate that could serve as a target for therapies designed to block downstream misfolding and aggregation events and thereby treat SOD1-associated ALS.
authors.affiliation: Department of Biochemistry, University of Toronto, Toronto Medical Discovery Tower 4-305, Toronto, ON, Canada M5G 1L7.
authors.forename: Vikram Khipple
authors.initials: VK
authors.lastname: Mulligan
authors.forename: Aaron
authors.initials: A
authors.lastname: Kerman
authors.forename: Rob C
authors.initials: RC
authors.lastname: Laister
authors.forename: Priya Roy
authors.initials: PR
authors.lastname: Sharda
authors.forename: Pharhad Eli
authors.initials: PE
authors.lastname: Arslan
authors.forename: Avijit
authors.initials: A
authors.lastname: Chakrabartty
mesh_terms: D000690:Amyotrophic Lateral Sclerosis / Q000503:physiopathology; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D008956:Models, Chemical; D008958:Models, Molecular; D010084:Oxidation-Reduction; D011489:Protein Denaturation; D017510:Protein Folding; D055503:Protein Multimerization; D013482:Superoxide Dismutase / Q000378:metabolism*; D000072105:Superoxide Dismutase-1
chemical_list: C000606290:SOD1 protein, human; D006861:Hydrogen Peroxide; D013482:Superoxide Dismutase; D000072105:Superoxide Dismutase-1