The lack of atomic structural information has become one of the major roadblocks in understanding ATP synthesis. This project is attempting to overcome this block through the elucidation -- by single crystal x-ray diffraction methods.-- of the structure of the F1-sector of the ATP synthase, the portion of the enzyme that contains all the catalytic and nucleotide binding sites. At the present level of resolution (3.6 Angstroms) density has been found for about 900 of the 1025 residues of the major subunits. This information has produced the most detailed description available of the quaternary structure of the F1-sector including the identification of alpha and beta subunits. Recent success in obtaining improved crystals has opened the door to extend the resolution of this study to 3.3 Angstroms. An electron density map at this higher resolution will resolve many of the present ambiguities in the chain connectivity; it will then be possible to complete tracing of the polypeptide chain and to refine the structure by crystallographic methods. The catalytic and nucleotide binding sites of the enzyme will be identified by inspection of the final structure and by studying complexes of the F1 with non-hydrolyzable ATP analogs using difference Fourier methods. This information will be used for evaluating existing mechanisms for ATP hydrolysis and synthesis and for proposing new mechanisms for experimental evaluation.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
2R01GM025432-12A2
Application #
3273004
Study Section
Biophysical Chemistry Study Section (BBCB)
Project Start
1978-07-01
Project End
1997-07-31
Budget Start
1993-08-01
Budget End
1994-07-31
Support Year
12
Fiscal Year
1993
Total Cost
Indirect Cost
Name
Johns Hopkins University
Department
Type
Schools of Medicine
DUNS #
045911138
City
Baltimore
State
MD
Country
United States
Zip Code
21218
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
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
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

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