|Strategies for stabilizing superoxide dismutase (SOD1), the protein destabilized in the most common form of familial amyotrophic lateral sclerosis.
|Year of Publication
|Auclair JR, Boggio KJ, Petsko GA, Ringe D, Agar JN
|Proc Natl Acad Sci U S A
|2010 Dec 14
|Amyotrophic 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
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.
|Proc. Natl. Acad. Sci. U.S.A.
|PubMed Central ID
|1R21NS071256 / NS / NINDS NIH HHS / United States