Strategies for stabilizing superoxide dismutase (SOD1), the protein destabilized in the most common form of familial amyotrophic lateral sclerosis.

TitleStrategies for stabilizing superoxide dismutase (SOD1), the protein destabilized in the most common form of familial amyotrophic lateral sclerosis.
Publication TypeJournal Article
Year of Publication2010
AuthorsAuclair JR, Boggio KJ, Petsko GA, Ringe D, Agar JN
JournalProc Natl Acad Sci U S A
Volume107
Issue50
Pagination21394-9
Date Published2010 Dec 14
ISSN1091-6490
KeywordsAmyotrophic Lateral Sclerosis, Animals, Cross-Linking Reagents, Disulfides, Enzyme Stability, Humans, Maleimides, Mice, Molecular Structure, Mutation, Protein Multimerization, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, Sulfhydryl Compounds, Superoxide Dismutase
Abstract

Amyotrophic lateral sclerosis (ALS) is a disorder characterized by the death of both upper and lower motor neurons and by 3- to 5-yr median survival postdiagnosis. The only US Food and Drug Administration-approved drug for the treatment of ALS, Riluzole, has at best, moderate effect on patient survival and quality of life; therefore innovative approaches are needed to combat neurodegenerative disease. Some familial forms of ALS (fALS) have been linked to mutations in the Cu/Zn superoxide dismutase (SOD1). The dominant inheritance of mutant SOD1 and lack of symptoms in knockout mice suggest a "gain of toxic function" as opposed to a loss of function. A prevailing hypothesis for the mechanism of the toxicity of fALS-SOD1 variants, or the gain of toxic function, involves dimer destabilization and dissociation as an early step in SOD1 aggregation. Therefore, stabilizing the SOD1 dimer, thus preventing aggregation, is a potential therapeutic strategy. Here, we report a strategy in which we chemically cross-link the SOD1 dimer using two adjacent cysteine residues on each respective monomer (Cys111). Stabilization, measured as an increase in melting temperature, of ∼20 °C and ∼45 °C was observed for two mutants, G93A and G85R, respectively. This stabilization is the largest for SOD1, and to the best of our knowledge, for any disease-related protein. In addition, chemical cross-linking conferred activity upon G85R, an otherwise inactive mutant. These results demonstrate that targeting these cysteine residues is an important new strategy for development of ALS therapies.

DOI10.1073/pnas.1015463107
Alternate JournalProc. Natl. Acad. Sci. U.S.A.
PubMed ID21098299
PubMed Central IDPMC3003092
Grant List1R21NS071256 / NS / NINDS NIH HHS / United States