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


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. Study holds ProTherm entries: 25853, 25854, 25855 Extra Details: presence of lower concentrations of cross-linker mass spectrometry; thiol-disulfide

Submission Details

ID: W87juAc24

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:56 p.m.

Version: 1

Publication Details
Auclair JR;Boggio KJ;Petsko GA;Ringe D;Agar JN,Proc. Natl. Acad. Sci. U.S.A. (2010) Strategies for stabilizing superoxide dismutase (SOD1), the protein destabilized in the most common form of familial amyotrophic lateral sclerosis. PMID:21098299
Additional Information

Structure view and single mutant data analysis

Study data

No weblogo for data of varying length.
Colors: D E R H K S T N Q A V I L M F Y W C G P
 

Data Distribution

Studies with similar sequences (approximate matches)

Correlation with other assays (exact sequence matches)


Relevant PDB Entries

Structure ID Release Date Resolution Structure Title
1F1G 2002-12-12 1.35 Crystal structure of yeast cuznsod exposed to nitric oxide
1EJ8 2000-04-05 1.55 CRYSTAL STRUCTURE OF DOMAIN 2 OF THE YEAST COPPER CHAPERONE FOR SUPEROXIDE DISMUTASE (LYS7) AT 1.55 A RESOLUTION
1JCV 1996-03-08 1.55 REDUCED BRIDGE-BROKEN YEAST CU/ZN SUPEROXIDE DISMUTASE LOW TEMPERATURE (-180C) STRUCTURE
1YAZ 2000-01-12 1.7 AZIDE-BOUND YEAST CU(II)/ZN SUPEROXIDE DISMUTASE ROOM TEMPERATURE (298K) STRUCTURE
2JCW 1999-06-08 1.7 REDUCED BRIDGE-BROKEN YEAST CU/ZN SUPEROXIDE DISMUTASE ROOM TEMPERATURE (298K) STRUCTURE
1F18 2002-12-18 1.7 Crystal structure of yeast copper-zinc superoxide dismutase mutant GLY85ARG
1YSO 1996-06-10 1.73 YEAST CU, ZN SUPEROXIDE DISMUTASE WITH THE REDUCED BRIDGE BROKEN
1F1A 2002-12-18 1.8 Crystal structure of yeast H48Q cuznsod fals mutant analog
1QUP 1999-12-10 1.8 CRYSTAL STRUCTURE OF THE COPPER CHAPERONE FOR SUPEROXIDE DISMUTASE
1B4T 1999-12-23 1.8 H48C YEAST CU(II)/ZN SUPEROXIDE DISMUTASE ROOM TEMPERATURE (298K) STRUCTURE
1B4L 1999-12-23 1.8 15 ATMOSPHERE OXYGEN YEAST CU/ZN SUPEROXIDE DISMUTASE ROOM TEMPERATURE (298K) STRUCTURE
1F1D 2002-12-18 2.1 Crystal structure of yeast H46C cuznsod mutant
5U9M 2017-05-31 2.35 Copper-Zinc Superoxide Dismutase is Activated through a Sulfenic Acid Intermediate at a Copper-ion Entry Site
1SDY 1994-01-31 2.5 STRUCTURE SOLUTION AND MOLECULAR DYNAMICS REFINEMENT OF THE YEAST CU,ZN ENZYME SUPEROXIDE DISMUTASE
1JK9 2001-09-05 2.9 Heterodimer between H48F-ySOD1 and yCCS

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
200.0 B,D Superoxide dismutase [Cu-Zn] P40202 CCS1_YEAST
198.6 A,C Superoxide dismutase [Cu-Zn] P00445 SODC_YEAST