title: Characterization of the binding of Fe(III) to F1ATPase from bovine heart mitochondria.
abstract: The binding Fe(III) to F1ATPase purified from beef heart mitochondria has been characterized by chemical analyses and EPR spectroscopy. F1ATPase binds 2 mol of Fe(III)/mol of protein selectively in the presence of saturating concentrations of ATP. In the absence of nucleotides or in the presence of either saturating ADP or limiting ATP concentrations, the enzyme binds 1 equivalent of Fe(III). F1ATPase pretreated with 5'-p- fluorosulfonylbenzoyladenosine, that selectively modifies the non-catalytic sites, binds only 1 mol of Fe(III)/mol of protein in the presence of either saturating ATP or ADP, Fe(III)-loaded F1ATPase containing either 1 or 2 equivalents of Fe(III) show identical EPR signals at g=4.3. The signals are not perturbed by the binding of nucleotides to the enzyme while they are altered by phosphate addition. These results indicate that F1ATPase contains two distinct Fe(III)-binding sites, which differ from nucleotide-binding sites, and that one of these sites is opened up for Fe(III) uptake by conformational changes induced by binding of ATP to the loose non-catalytic site.
authors.affiliation: Department of Biomedical Sciences and Technologies, University of Udine, Italy.
authors.forename: G
authors.initials: G
authors.lastname: Lippe
authors.forename: F
authors.initials: F
authors.lastname: Polizio
authors.forename: F
authors.initials: F
authors.lastname: Di Pancrazio
authors.forename: F
authors.initials: F
authors.lastname: Dabbeni-Sala
authors.forename: N
authors.initials: N
authors.lastname: Bortolotti
authors.forename: A
authors.initials: A
authors.lastname: Desideri
authors.forename: I
authors.initials: I
authors.lastname: Mavelli
mesh_terms: D000244:Adenosine Diphosphate / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D001665:Binding Sites; D002417:Cattle; D004578:Electron Spin Resonance Spectroscopy; D005290:Ferric Compounds / Q000032:analysis / Q000378:metabolism*; D006868:Hydrolysis; D008929:Mitochondria, Heart / Q000201:enzymology*; D006180:Proton-Translocating ATPases / Q000032:analysis / Q000378:metabolism*
chemical_list: D005290:Ferric Compounds; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D006180:Proton-Translocating ATPases