title: Comparison between single-molecule and X-ray crystallography data on yeast F1-ATPase.
abstract: Single molecule studies in recent decades have elucidated the full chemo-mechanical cycle of F1-ATPase, mostly based on F1 from thermophilic bacteria. In contrast, high-resolution crystal structures are only available for mitochondrial F1. Here we present high resolution single molecule rotational data on F1 from Saccharomyces cerevisiae, obtained using new high throughput detection and analysis tools. Rotational data are presented for the wild type mitochondrial enzyme, a "liver" isoform, and six mutant forms of yeast F1 that have previously been demonstrated to be less efficient or partially uncoupled. The wild-type and "liver" isoforms show the same qualitative features as F1 from Escherichia coli and thermophilic bacteria. The analysis of the mutant forms revealed a delay at the catalytic dwell and associated decrease in Vmax, with magnitudes consistent with the level of disruption seen in the crystal structures. At least one of the mutant forms shows a previously un-observed dwell at the ATP binding angle, potentially attributable to slowed release of ADP. We discuss the correlation between crystal structures and single molecule results.
authors.affiliation: Department of Physics, University of Oxford, Clarendon Laboratory, Parks Road, Oxford, OX1 3PU UK.
authors.forename: Bradley C
authors.initials: BC
authors.lastname: Steel
authors.affiliation: 1] Department of Physics, University of Oxford, Clarendon Laboratory, Parks Road, Oxford, OX1 3PU UK [2] Centre de Biochimie Structurale, 29 Rue de Navacelles, Montpellier, 34000, France.
authors.forename: Ashley L
authors.initials: AL
authors.lastname: Nord
authors.affiliation: Department of Biochemistry and Molecular Biology, Rosalind Franklin University of Medicine and Science, The Chicago Medical School, North Chicago, Illinois, 60064, USA.
authors.forename: Yamin
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Department of Biochemistry and Molecular Biology, Rosalind Franklin University of Medicine and Science, The Chicago Medical School, North Chicago, Illinois, 60064, USA.
authors.forename: Vijayakanth
authors.initials: V
authors.lastname: Pagadala
authors.affiliation: Department of Biochemistry and Molecular Biology, Rosalind Franklin University of Medicine and Science, The Chicago Medical School, North Chicago, Illinois, 60064, USA.
authors.forename: David M
authors.initials: DM
authors.lastname: Mueller
authors.affiliation: Department of Physics, University of Oxford, Clarendon Laboratory, Parks Road, Oxford, OX1 3PU UK.
authors.forename: Richard M
authors.initials: RM
authors.lastname: Berry
mesh_terms: D018360:Crystallography, X-Ray; D007527:Isoenzymes; D007700:Kinetics; D008958:Models, Molecular; D009154:Mutation; D011487:Protein Conformation; D006180:Proton-Translocating ATPases / Q000737:chemistry* / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000737:chemistry* / Q000201:enzymology / Q000235:genetics
chemical_list: D007527:Isoenzymes; D006180:Proton-Translocating ATPases