title: Human superoxide dismutase 1 (hSOD1) maturation through interaction with human copper chaperone for SOD1 (hCCS).
abstract: Copper chaperone for superoxide dismutase 1 (SOD1), CCS, is the physiological partner for the complex mechanism of SOD1 maturation. We report an in vitro model for human CCS-dependent SOD1 maturation based on the study of the interactions of human SOD1 (hSOD1) with full-length WT human CCS (hCCS), as well as with hCCS mutants and various truncated constructs comprising one or two of the protein's three domains. The synergy between electrospray ionization mass spectrometry (ESI-MS) and NMR is fully exploited. This is an in vitro study of this process at the molecular level. Domain 1 of hCCS is necessary to load hSOD1 with Cu(I), requiring the heterodimeric complex formation with hSOD1 fostered by the interaction with domain 2. Domain 3 is responsible for the catalytic formation of the hSOD1 Cys-57-Cys-146 disulfide bond, which involves both hCCS Cys-244 and Cys-246 via disulfide transfer.
authors.affiliation: Magnetic Resonance Center, University of Florence, 50019 Sesto Fiorentino, Italy. banci@cerm.unifi.it
authors.forename: Lucia
authors.initials: L
authors.lastname: Banci
authors.forename: Ivano
authors.initials: I
authors.lastname: Bertini
authors.forename: Francesca
authors.initials: F
authors.lastname: Cantini
authors.forename: Tatiana
authors.initials: T
authors.lastname: Kozyreva
authors.forename: Chiara
authors.initials: C
authors.lastname: Massagni
authors.forename: Peep
authors.initials: P
authors.lastname: Palumaa
authors.forename: Jeffrey T
authors.initials: JT
authors.lastname: Rubino
authors.forename: Kairit
authors.initials: K
authors.lastname: Zovo
mesh_terms: D001665:Binding Sites; D003300:Copper / Q000737:chemistry*; D003545:Cysteine / Q000737:chemistry; D004220:Disulfides / Q000737:chemistry; D006801:Humans; D007700:Kinetics; D008190:Lyases / Q000737:chemistry / Q000502:physiology*; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D018832:Molecular Chaperones / Q000378:metabolism; D009154:Mutation; D010084:Oxidation-Reduction; D011485:Protein Binding; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D013482:Superoxide Dismutase / Q000235:genetics* / Q000502:physiology*; D000072105:Superoxide Dismutase-1; D013997:Time Factors
chemical_list: D004220:Disulfides; D018832:Molecular Chaperones; C000606290:SOD1 protein, human; D003300:Copper; D013482:Superoxide Dismutase; D000072105:Superoxide Dismutase-1; D008190:Lyases; C031581:cytochrome C synthetase; D003545:Cysteine