title: A pivotal role of Zn-binding residues in the function of the copper chaperone for SOD1.
abstract: A Cu chaperone for SOD1 (CCS) is required for the incorporation of copper ion into the protein. To investigate the roles of the conserved metal-binding residues in CCS, we introduced amino acid substitutions into human CCS and examined the function of the mutant CCS by transforming a mutant yeast strain, SY2950, which lacks the lys7 gene, a yeast orthologue of the mammalian CCS. Mutant CCS in which amino acid residues His147 and Asp167 were substituted by Ala exhibited a decreased ability to complement the growth of SY2950 under Lys-deficient conditions. This is because the mutations made the human CCS function in a less efficient manner, especially under metal-restricted conditions, leaving Cu,Zn-SOD in an apo-form. Since the His and Asp residues are both responsible for binding Zn which would serve to maintain the folded structure, the structural integrity supported by the coordinated Zn ion would be essential for CCS function.
authors.affiliation: Department of Biochemistry, Osaka University Medical School, 2-2 Yamadaoka, Suita, Osaka, 565-0871, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Endo
authors.forename: T
authors.initials: T
authors.lastname: Fujii
authors.forename: K
authors.initials: K
authors.lastname: Sato
authors.forename: N
authors.initials: N
authors.lastname: Taniguchi
authors.forename: J
authors.initials: J
authors.lastname: Fujii
mesh_terms: D000595:Amino Acid Sequence; D019943:Amino Acid Substitution / Q000235:genetics; D001059:Apoproteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D001665:Binding Sites; D005656:Fungal Proteins / Q000235:genetics; D017353:Gene Deletion; D005800:Genes, Fungal / Q000235:genetics; D005816:Genetic Complementation Test; D006801:Humans; D018832:Molecular Chaperones / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D021381:Protein Transport; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; D016415:Sequence Alignment; D013482:Superoxide Dismutase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D015003:Yeasts / Q000201:enzymology / Q000235:genetics / Q000254:growth & development; D015032:Zinc / Q000378:metabolism*
chemical_list: D001059:Apoproteins; C114678:CCS protein, human; C491960:CCS1 protein, S cerevisiae; D005656:Fungal Proteins; D018832:Molecular Chaperones; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; C095735:LYS7 protein, S cerevisiae; D013482:Superoxide Dismutase; D015032:Zinc