title: Oxidation of the tryptophan 32 residue of human superoxide dismutase 1 caused by its bicarbonate-dependent peroxidase activity triggers the non-amyloid aggregation of the enzyme.
abstract: The role of oxidative post-translational modifications of human superoxide dismutase 1 (hSOD1) in the amyotrophic lateral sclerosis (ALS) pathology is an attractive hypothesis to explore based on several lines of evidence. Among them, the remarkable stability of hSOD1(WT) and several of its ALS-associated mutants suggests that hSOD1 oxidation may precede its conversion to the unfolded and aggregated forms found in ALS patients. The bicarbonate-dependent peroxidase activity of hSOD1 causes oxidation of its own solvent-exposed Trp(32) residue. The resulting products are apparently different from those produced in the absence of bicarbonate and are most likely specific for simian SOD1s, which contain the Trp(32) residue. The aims of this work were to examine whether the bicarbonate-dependent peroxidase activity of hSOD1 (hSOD1(WT) and hSOD1(G93A) mutant) triggers aggregation of the enzyme and to comprehend the role of the Trp(32) residue in the process. The results showed that Trp(32) residues of both enzymes are oxidized to a similar extent to hSOD1-derived tryptophanyl radicals. These radicals decayed to hSOD1-N-formylkynurenine and hSOD1-kynurenine or to a hSOD1 covalent dimer cross-linked by a ditryptophan bond, causing hSOD1 unfolding, oligomerization, and non-amyloid aggregation. The latter process was inhibited by tempol, which recombines with the hSOD1-derived tryptophanyl radical, and did not occur in the absence of bicarbonate or with enzymes that lack the Trp(32) residue (bovine SOD1 and hSOD1(W32F) mutant). The results support a role for the oxidation products of the hSOD1-Trp(32) residue, particularly the covalent dimer, in triggering the non-amyloid aggregation of hSOD1.
authors.affiliation: Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, 05513-970 São Paulo, SP, Brazil.
authors.forename: Fernando R
authors.initials: FR
authors.lastname: Coelho
authors.affiliation: Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, 05513-970 São Paulo, SP, Brazil.
authors.forename: Asif
authors.initials: A
authors.lastname: Iqbal
authors.affiliation: Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, 05513-970 São Paulo, SP, Brazil.
authors.forename: Edlaine
authors.initials: E
authors.lastname: Linares
authors.affiliation: Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, 05513-970 São Paulo, SP, Brazil.
authors.forename: Daniel F
authors.initials: DF
authors.lastname: Silva
authors.affiliation: Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, 05513-970 São Paulo, SP, Brazil.
authors.forename: Filipe S
authors.initials: FS
authors.lastname: Lima
authors.affiliation: Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, 05513-970 São Paulo, SP, Brazil.
authors.forename: Iolanda M
authors.initials: IM
authors.lastname: Cuccovia
authors.affiliation: Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, 05513-970 São Paulo, SP, Brazil. Electronic address: oaugusto@iq.usp.br.
authors.forename: Ohara
authors.initials: O
authors.lastname: Augusto
mesh_terms: D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D001639:Bicarbonates / Q000737:chemistry; D006801:Humans; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D066263:Protein Aggregation, Pathological; D050050:Protein Carbonylation; D055503:Protein Multimerization; D013482:Superoxide Dismutase / Q000737:chemistry* / Q000235:genetics; D000072105:Superoxide Dismutase-1; D014364:Tryptophan / Q000737:chemistry*
chemical_list: D001639:Bicarbonates; C000606290:SOD1 protein, human; D014364:Tryptophan; D013482:Superoxide Dismutase; D000072105:Superoxide Dismutase-1
keywords: Amyotrophic Lateral Sclerosis (ALS) (Lou Gehrig Disease); Bicarbonate; Bicarbonate-dependent Peroxidase Activity; Ditryptophan; Free Radicals; Human SOD1; Protein Aggregation; Protein Oxidation; Superoxide Dismutase (SOD)