title: Altered thiol chemistry in human amyotrophic lateral sclerosis-linked mutants of superoxide dismutase 1.
abstract: Neurodegenerative diseases share a common characteristic, the presence of intracellular or extracellular deposits of protein aggregates in nervous tissues. Amyotrophic Lateral Sclerosis (ALS) is a severe and fatal neurodegenerative disorder, which affects preferentially motoneurons. Changes in the redox state of superoxide dismutase 1 (SOD1) are associated with the onset and development of familial forms of ALS. In human SOD1 (hSOD1), a conserved disulfide bond and two free cysteine residues can engage in anomalous thiol/disulfide exchange resulting in non-native disulfides, a hallmark of ALS that is related to protein misfolding and aggregation. Because of the many competing reaction pathways, traditional bulk techniques fall short at quantifying individual thiol/disulfide exchange reactions. Here, we adapt recently developed single-bond chemistry techniques to study individual disulfide isomerization reactions in hSOD1. Mechanical unfolding of hSOD1 leads to the formation of a polypeptide loop held by the disulfide. This loop behaves as a molecular jump rope that brings reactive Cys-111 close to the disulfide. Using force-clamp spectroscopy, we monitor nucleophilic attack of Cys-111 at either sulfur of the disulfide and determine the selectivity of the reaction. Disease-causing mutations G93A and A4V show greatly altered reactivity patterns, which may contribute to the progression of familial ALS.
authors.affiliation: Laboratory of Cellular and Molecular Neurobiology, Department of Pathology and Experimental Therapeutics, Faculty of Medicine-Campus Bellvitge, University of Barcelona, Feixa Llarga s/n. Hospitalet de Llobregat, 08907 Barcelona, Spain,; Bellvitge Biomedical Research Institute (IDIBELL), Gran Via de l'Hospitalet, 199-203, L'Hospitalet de Llobregat, Barcelona, 08908 Barcelona, Spain,. Electronic address: solsona@ub.edu.
authors.forename: Carles
authors.initials: C
authors.lastname: Solsona
authors.affiliation: Department of Biochemistry and Molecular Biophysics, Columbia University Medical Center, New York, New York 10032,; Department of Biological Sciences, Columbia University, New York, New York 10027, and.
authors.forename: Thomas B
authors.initials: TB
authors.lastname: Kahn
authors.affiliation: Department of Biological Sciences, Columbia University, New York, New York 10027, and.
authors.forename: Carmen L
authors.initials: CL
authors.lastname: Badilla
authors.affiliation: Laboratory of Cellular and Molecular Neurobiology, Department of Pathology and Experimental Therapeutics, Faculty of Medicine-Campus Bellvitge, University of Barcelona, Feixa Llarga s/n. Hospitalet de Llobregat, 08907 Barcelona, Spain,; Bellvitge Biomedical Research Institute (IDIBELL), Gran Via de l'Hospitalet, 199-203, L'Hospitalet de Llobregat, Barcelona, 08908 Barcelona, Spain.
authors.forename: Cristina
authors.initials: C
authors.lastname: Álvarez-Zaldiernas
authors.affiliation: Laboratory of Cellular and Molecular Neurobiology, Department of Pathology and Experimental Therapeutics, Faculty of Medicine-Campus Bellvitge, University of Barcelona, Feixa Llarga s/n. Hospitalet de Llobregat, 08907 Barcelona, Spain,; Bellvitge Biomedical Research Institute (IDIBELL), Gran Via de l'Hospitalet, 199-203, L'Hospitalet de Llobregat, Barcelona, 08908 Barcelona, Spain.
authors.forename: Juan
authors.initials: J
authors.lastname: Blasi
authors.affiliation: Department of Biological Sciences, Columbia University, New York, New York 10027, and.
authors.forename: Julio M
authors.initials: JM
authors.lastname: Fernandez
authors.affiliation: Department of Biological Sciences, Columbia University, New York, New York 10027, and; Vascular Biology and Inflammation Department, Centro Nacional de Investigaciones Cardiovasculares (CNIC), Cl. Melchor Fernández Almagro 3, 28029 Madrid, Spain.
authors.forename: Jorge
authors.initials: J
authors.lastname: Alegre-Cebollada
mesh_terms: D019943:Amino Acid Substitution; D000690:Amyotrophic Lateral Sclerosis / Q000201:enzymology* / Q000235:genetics; D003545:Cysteine / Q000737:chemistry* / Q000235:genetics; D004220:Disulfides / Q000737:chemistry*; D006801:Humans; D020125:Mutation, Missense; D010084:Oxidation-Reduction; D017433:Protein Structure, Secondary; D058767:Protein Unfolding; D013482:Superoxide Dismutase / Q000737:chemistry* / Q000235:genetics; D000072105:Superoxide Dismutase-1
chemical_list: D004220:Disulfides; C000606290:SOD1 protein, human; D013482:Superoxide Dismutase; D000072105:Superoxide Dismutase-1; D003545:Cysteine
keywords: Amyotrophic Lateral Sclerosis (ALS) (Lou Gehrig disease); Atomic Force Microscopy (AFM); Atomic Force Spectroscopy; Cysteine-mediated Cross-linking; Disulfide; Neurodegenerative Disease; Protein Misfolding; Superoxide Dismutase (SOD)
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