title: Structure of the two transmembrane Cu+ transport sites of the Cu+ -ATPases.
abstract: Cu(+)-ATPases drive metal efflux from the cell cytoplasm. Paramount to this function is the binding of Cu(+) within the transmembrane region and its coupled translocation across the permeability barrier. Here, we describe the two transmembrane Cu(+) transport sites present in Archaeoglobus fulgidus CopA. Both sites can be independently loaded with Cu(+). However, their simultaneous occupation is associated with enzyme turnover. Site I is constituted by two Cys in transmembrane segment (TM) 6 and a Tyr in TM7. An Asn in TM7 and Met and Ser in TM8 form Site II. Single site x-ray spectroscopic analysis indicates a trigonal coordination in both sites. This architecture is distinct from that observed in Cu(+)-trafficking chaperones and classical cuproproteins. The high affinity of these sites for Cu(+) (Site I K(a)=1.3 fM(-1), Site II K(a)=1.1 fM(-1)), in conjunction with reversible direct Cu(+) transfer from chaperones, points to a transport mechanism where backward release of free Cu(+) to the cytoplasm is largely prevented.
authors.affiliation: Department of Chemistry and Biochemistry, Worcester Polytechnic Institute, Worcester, Massachusetts 01609, USA.
authors.forename: Manuel
authors.initials: M
authors.lastname: González-Guerrero
authors.forename: Elif
authors.initials: E
authors.lastname: Eren
authors.forename: Swati
authors.initials: S
authors.lastname: Rawat
authors.forename: Timothy L
authors.initials: TL
authors.lastname: Stemmler
authors.forename: José M
authors.initials: JM
authors.lastname: Argüello
mesh_terms: D000251:Adenosine Triphosphatases / Q000737:chemistry*; D019611:Archaeoglobus fulgidus / Q000378:metabolism*; D001665:Binding Sites; D001692:Biological Transport; D003300:Copper / Q000737:chemistry*; D003545:Cysteine / Q000737:chemistry; D006639:Histidine / Q000737:chemistry; D007700:Kinetics; D008954:Models, Biological; D018832:Molecular Chaperones / Q000737:chemistry; D016297:Mutagenesis, Site-Directed; D009596:Nitrophenols / Q000737:chemistry; D009943:Organophosphorus Compounds / Q000737:chemistry; D021381:Protein Transport; D014443:Tyrosine / Q000737:chemistry; D014965:X-Rays
chemical_list: D018832:Molecular Chaperones; D009596:Nitrophenols; D009943:Organophosphorus Compounds; C008644:nitrophenylphosphate; D014443:Tyrosine; D006639:Histidine; D003300:Copper; D000251:Adenosine Triphosphatases; D003545:Cysteine
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