title: The nucleotide-independent Fe(III)-binding site is located on beta subunit of the mitochondrial F(1)-ATPase.
abstract: Upon separation of a crude preparation of beta subunit ("beta fraction") from mitochondrial F(1)-ATPase containing one equivalent of Fe(III) in the nucleotide-independent site (1Fe(III)-loaded MF(1)), Fe(III) is almost completely recovered. CD spectra show that "beta fraction" maintains the structural changes induced by Fe(III) in the whole enzyme. In accordance, EPR reveals that the Fe(III) site geometry is conserved in "beta fraction." Moreover, the EPR spectra of 1Fe(III)-loaded MF(1) and its "beta fraction" undergo similar changes of the line-shape upon Pi binding at the catalytic site, indicating that the Pi and Fe(III) are proximal on beta. Highly purified beta in nucleotide-free form binds 1mol of Fe(III)/mol of protein. MF(1) "freezed" by inhibitors with two beta in closed conformation and one beta in open or half-closed conformation binds 1mol of Fe(III)/mol of enzyme. Therefore, the Fe(III) site location in the unique beta subunit not adopting the closed conformation is proposed.
authors.affiliation: Department of Biomedical Sciences and Technologies, University of Udine, P.le Kolbe 4, 33100 Udine, Italy. glippe@makek.dstb.uniud.it
authors.forename: G
authors.initials: G
authors.lastname: Lippe
authors.forename: F
authors.initials: F
authors.lastname: Di Pancrazio
authors.forename: S
authors.initials: S
authors.lastname: Contessi
authors.forename: N
authors.initials: N
authors.lastname: Bortolotti
authors.forename: F
authors.initials: F
authors.lastname: Polizio
authors.forename: I
authors.initials: I
authors.lastname: Mavelli
mesh_terms: D000818:Animals; D001665:Binding Sites; D002417:Cattle; D002942:Circular Dichroism; D004578:Electron Spin Resonance Spectroscopy; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005290:Ferric Compounds / Q000378:metabolism*; D008929:Mitochondria, Heart / Q000201:enzymology*; D021122:Protein Subunits; D006180:Proton-Translocating ATPases / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000378:metabolism
chemical_list: D004791:Enzyme Inhibitors; D005290:Ferric Compounds; D021122:Protein Subunits; D006180:Proton-Translocating ATPases