title: F1F0-ATP synthase: development of direct optical probes of the catalytic mechanism.
abstract: Using strategically-placed tryptophan (Trp) residues as optical probes to monitor nucleotide binding and hydrolysis, we demonstrate that all three catalytic nucleotide binding sites in F1-ATPase must be filled to obtain physiological (Vmax) MgATP hydrolysis rates. At Vmax hydrolysis rates, the predominant enzyme species has one of the three catalytic sites filled with unhydrolyzed substrate MgATP, the other two sites are filled with product MgADP. A specifically-inserted Trp probe was also developed to characterize nucleotide binding to the noncatalytic sites, and a model to explain the specificity of these sites is shown. These sites appear to play no role in ATP hydrolysis.
authors.affiliation: University of Rochester Medical Center, Department of Biochemistry, NY 14642, USA. weber@medinfo.rochester.edu
authors.forename: J
authors.initials: J
authors.lastname: Weber
authors.forename: A E
authors.initials: AE
authors.lastname: Senior
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D001665:Binding Sites; D006868:Hydrolysis; D009711:Nucleotides / Q000378:metabolism; D006180:Proton-Translocating ATPases / Q000737:chemistry / Q000378:metabolism*
chemical_list: D009711:Nucleotides; D000255:Adenosine Triphosphate; D006180:Proton-Translocating ATPases