title: Structure of Cu(I)-bound DJ-1 reveals a biscysteinate metal binding site at the homodimer interface: insights into mutational inactivation of DJ-1 in Parkinsonism.
abstract: The Parkinsonism-associated protein DJ-1 has been suggested to activate the Cu-Zn superoxide dismutase (SOD1) by providing its copper cofactor. The structural and chemical means by which DJ-1 could support this function is unknown. In this study, we characterize the molecular interaction of DJ-1 with Cu(I). Mass spectrometric analysis indicates binding of one Cu(I) ion per DJ-1 homodimer. The crystal structure of DJ-1 bound to Cu(I) confirms metal coordination through a docking accessible biscysteinate site formed by juxtaposed cysteine residues at the homodimer interface. Spectroscopy in crystallo validates the identity and oxidation state of the bound metal. The measured subfemtomolar dissociation constant (Kd = 6.41 × 10(-16) M) of DJ-1 for Cu(I) supports the physiological retention of the metal ion. Our results highlight the requirement of a stable homodimer for copper binding by DJ-1. Parkinsonism-linked mutations that weaken homodimer interactions will compromise this capability.
authors.affiliation: Department of Molecular and Applied Biosciences, University of Westminster , 115 New Cavendish Street, London W1W 6UW, United Kingdom.
authors.forename: M Rhyan
authors.initials: MR
authors.lastname: Puno
authors.forename: Nisha A
authors.initials: NA
authors.lastname: Patel
authors.forename: Simon Geir
authors.initials: SG
authors.lastname: Møller
authors.forename: Carol V
authors.initials: CV
authors.lastname: Robinson
authors.forename: Peter C E
authors.initials: PC
authors.lastname: Moody
authors.forename: Mark
authors.initials: M
authors.lastname: Odell
mesh_terms: D001665:Binding Sites; D003300:Copper / Q000737:chemistry* / Q000378:metabolism; D018360:Crystallography, X-Ray; D003545:Cysteine / Q000737:chemistry*; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D013058:Mass Spectrometry; D008670:Metals / Q000737:chemistry; D008958:Models, Molecular; D009154:Mutation / Q000502:physiology*; D015513:Oncogene Proteins / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D010300:Parkinson Disease / Q000235:genetics* / Q000378:metabolism*; D011487:Protein Conformation; D000071617:Protein Deglycase DJ-1; D021241:Spectrometry, Mass, Electrospray Ionization; D013482:Superoxide Dismutase / Q000737:chemistry; D000072105:Superoxide Dismutase-1
chemical_list: D047908:Intracellular Signaling Peptides and Proteins; D008670:Metals; D015513:Oncogene Proteins; C000606290:SOD1 protein, human; D003300:Copper; D013482:Superoxide Dismutase; D000072105:Superoxide Dismutase-1; C105131:PARK7 protein, human; D000071617:Protein Deglycase DJ-1; D003545:Cysteine