title: Cu/Zn superoxide dismutase mutants associated with amyotrophic lateral sclerosis show enhanced formation of aggregates in vitro.
abstract: Mutations in Cu/Zn superoxide dismutase (SOD) are associated with the fatal neurodegenerative disorder amyotrophic lateral sclerosis (ALS). There is considerable evidence that mutant SOD has a gain of toxic function; however, the mechanism of this toxicity is not known. We report here that purified SOD forms aggregates in vitro under destabilizing solution conditions by a process involving a transition from small amorphous species to fibrils. The assembly process and the tinctorial and structural properties of the in vitro aggregates resemble those for aggregates observed in vivo. Furthermore, the familial ALS SOD mutations A4V, G93A, G93R, and E100G decrease protein stability, which correlates with an increase in the propensity of the mutants to form aggregates. These mutations also increase the rate of protein unfolding. Our results suggest three possible mechanisms for the increase in aggregation: (i) an increase in the equilibrium population of unfolded or of partially unfolded states, (ii) an increase in the rate of unfolding, and (iii) a decrease in the rate of folding. Our data support the hypothesis that the gain of toxic function for many different familial ALS-associated mutant SODs is a consequence of protein destabilization, which leads to an increase in the formation of cytotoxic protein aggregates.
authors.affiliation: Guelph-Waterloo Centre for Graduate Studies in Chemistry and Biochemistry, University of Waterloo, Waterloo, ON, Canada N2L 3G1.
authors.forename: P B
authors.initials: PB
authors.lastname: Stathopulos
authors.forename: J A O
authors.initials: JA
authors.lastname: Rumfeldt
authors.forename: G A
authors.initials: GA
authors.lastname: Scholz
authors.forename: R A
authors.initials: RA
authors.lastname: Irani
authors.forename: H E
authors.initials: HE
authors.lastname: Frey
authors.forename: R A
authors.initials: RA
authors.lastname: Hallewell
authors.forename: J R
authors.initials: JR
authors.lastname: Lepock
authors.forename: E M
authors.initials: EM
authors.lastname: Meiering
mesh_terms: D000690:Amyotrophic Lateral Sclerosis / Q000201:enzymology* / Q000235:genetics*; D001051:Apoenzymes / Q000737:chemistry / Q000235:genetics; D004795:Enzyme Stability / Q000235:genetics; D020035:Holoenzymes / Q000737:chemistry / Q000235:genetics; D006358:Hot Temperature; D006801:Humans; D066298:In Vitro Techniques; D046911:Macromolecular Substances; D008854:Microscopy, Electron; D009154:Mutation; D017510:Protein Folding; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D013482:Superoxide Dismutase / Q000737:chemistry* / Q000235:genetics* / Q000648:ultrastructure; D014270:Trifluoroethanol
chemical_list: D001051:Apoenzymes; D020035:Holoenzymes; D046911:Macromolecular Substances; D011994:Recombinant Proteins; D014270:Trifluoroethanol; D013482:Superoxide Dismutase