Abstract

The mechanism of oxidative phosphorylation was and has remained one of the most challenging problems in biochemistry. Central to the elucidation of the mechanism of this pathway is a complete understanding of the functional and structural properties of mitochondrial ATPase (ATP synthetase), and enzyme system that catalyses the last of the oxidative phosphorylation reactions. Crystals of the soluble portion of this enzyme (F1-ATPase) were obtained in our laboratory and the structure is being studied at 9 A resolution. In this project we propose to continue our single crystal X-ray diffraction studies of the determination of the three-dimensional structure of the F1-sector of the ATP synthetase from rat liver mitochondria. Our present results will be extended through the inclusion of diffraction data to 3.5 A resolution. The structures of the complexes of F1-ATPase with substrates, products and effectors (or their analogs) will also be determined. The determination of these structures will address in a very direct way some of the outstanding questions in oxidative phosphorylation and provide invaluable information for the understanding of ATP-dependent ion translocation processes.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM025432-07
Application #
3273005
Study Section
Biophysics and Biophysical Chemistry B Study Section (BBCB)
Project Start
1978-07-01
Project End
1986-03-31
Budget Start
1985-04-01
Budget End
1986-03-31
Support Year
7
Fiscal Year
1985
Total Cost
Indirect Cost

  Institution

Name
Johns Hopkins University
Department
Type
Schools of Medicine
DUNS #
045911138
City
Baltimore
State
MD
Country
United States
Zip Code
21218

  Publications

Bianchet, M A; Pedersen, P L; Amzel, L M (2000) Notes on the mechanism of ATP synthesis. J Bioenerg Biomembr 32:517-21
Bianchet, M A; Hullihen, J; Pedersen, P L et al. (1998) The 2.8-A structure of rat liver F1-ATPase: configuration of a critical intermediate in ATP synthesis/hydrolysis. Proc Natl Acad Sci U S A 95:11065-70
Bianchet, M A; Ko, Y H; Amzel, L M et al. (1997) Modeling of nucleotide binding domains of ABC transporter proteins based on a F1-ATPase/recA topology: structural model of the nucleotide binding domains of the cystic fibrosis transmembrane conductance regulator (CFTR). J Bioenerg Biomembr 29:503-24
Pedersen, P L; Hullihen, J; Bianchet, M et al. (1995) Rat liver ATP synthase. Relationship of the unique substructure of the F1 moiety to its nucleotide binding properties, enzymatic states, and crystalline form. J Biol Chem 270:1775-84
Pedersen, P L; Amzel, L M (1993) ATP synthases. Structure, reaction center, mechanism, and regulation of one of nature's most unique machines. J Biol Chem 268:9937-40
Thomas, P J; Bianchet, M; Garboczi, D N et al. (1992) ATP synthase: structure-function relationships. Biochim Biophys Acta 1101:228-31
Pedersen, P L; Thomas, P J; Garboczi, D N et al. (1992) F-type ATPases: are nucleotide domains in adenylate kinase appropriate models for nucleotide domains in ATP synthase/ATPase complexes? Ann N Y Acad Sci 671:359-65
Amzel, L M; Bianchet, M A; Pedersen, P L (1992) Quaternary structure of ATP synthases: symmetry and asymmetry in the F1 moiety. J Bioenerg Biomembr 24:429-33
Bianchet, M; Ysern, X; Hullihen, J et al. (1991) Mitochondrial ATP synthase. Quaternary structure of the F1 moiety at 3.6 A determined by x-ray diffraction analysis. J Biol Chem 266:21197-201
Hurley, T D; Bosron, W F; Hamilton, J A et al. (1991) Structure of human beta 1 beta 1 alcohol dehydrogenase: catalytic effects of non-active-site substitutions. Proc Natl Acad Sci U S A 88:8149-53

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