title: Mechanistic aspects of hSOD1 maturation from the solution structure of Cu(I) -loaded hCCS domain 1 and analysis of disulfide-free hSOD1 mutants.
abstract: Superoxide dismutase 1 (SOD1) maturation within the cell is mainly accomplished with the SOD1-specific chaperone, CCS, a dimeric protein with three distinct domains in each monomer. We recently showed that the first domain of human CCS (hCCSD1) is responsible for copper transfer to its protein partner, human SOD1 (hSOD1). The NMR solution structure of the copper(I)-loaded form of hCCSD1 reported here contributes further to characterization of the copper-transfer mechanism to hSOD1. NMR spectroscopy was also used to examine the hSOD1 mutants C57A, C146A, and C57A/C146A, which are unable to form the structurally conserved disulfide bond in SOD1, in order to investigate the role of these cysteines during hSOD1 copper acquisition. Together, the information on both hCCS and hSOD1, along with a sequence analysis of eukaryotic CCSD1, allows us to propose important mechanistic aspects regarding the copper-transfer process from hCCS to hSOD1.
authors.affiliation: Magnetic Resonance Center (CERM), University of Florence, Via L. Sacconi 6, 50019 Sesto Fiorentino (Italy); Department of Chemistry, University of Florence, Via della Lastruccia 3, 50019 Sesto Fiorentino (Italy); Fondazione Farmacogenomica FiorGen onlus Via L. Sacconi 6, 50019, Sesto Fiorentino (Italy). banci@cerm.unifi.it.
authors.forename: Lucia
authors.initials: L
authors.lastname: Banci
authors.forename: Francesca
authors.initials: F
authors.lastname: Cantini
authors.forename: Tatiana
authors.initials: T
authors.lastname: Kozyreva
authors.forename: Jeffrey T
authors.initials: JT
authors.lastname: Rubino
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D003300:Copper / Q000737:chemistry* / Q000378:metabolism; D004220:Disulfides / Q000737:chemistry*; D006801:Humans; D018832:Molecular Chaperones / Q000737:chemistry* / Q000378:metabolism; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D019906:Nuclear Magnetic Resonance, Biomolecular; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D016415:Sequence Alignment; D013482:Superoxide Dismutase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D000072105:Superoxide Dismutase-1
chemical_list: D004220:Disulfides; D018832:Molecular Chaperones; D011994:Recombinant Proteins; C000606290:SOD1 protein, human; D003300:Copper; D013482:Superoxide Dismutase; D000072105:Superoxide Dismutase-1
keywords: CCS; NMR spectroscopy; SOD1; chaperone proteins; copper transport