The mitochondrial F1Fo ATP synthase is responsible for the synthesis 90% of the ATP under aerobic conditions. The ATP synthase is a multimeric protein complex with an overall molecular weight greater than 550,000 Da. A portion of the ATP synthase is embedded in the mitochondrial membrane and acts as a proton turbine and a portion is in the matrix space and acts as a rotary engine that phosphorylates ADP. In the prior award period, we have solved the high resolution structures of 4 mutant forms of the F1 ATPase and determined the structural basis of mutations in F1 that uncouple proton translocation from ATP synthesis. The coupling of the ATP synthase is key to energy metabolism and yet the mechanism is poorly understood. We have also determined the high-resolution structures of the yeast c10-ring at pH 5.5, 6.1, 8.3, modified with DCCD, and bound with oligomycin. The current project builds upon our advances and expands our goals.
One aim of this project is to determine the high-resolution crystal structure of the ATP synthase complex from yeast S. cerevisiae.
This aim will give critical structural details into the understanding of the mechanism of proton translocation coupled to ATP synthesis. A number of genetic modifications will be used to facilitate crystallization of the entire complex. The high-resolution structure of the ATP synthase will provide the critical features to understand and treat diseases such as cancer and diabetes.

Public Health Relevance

The mitochondrial ATP synthase is an essential enzyme and involved in the energy conversion from carbohydrates, fats, and protein, to ATP, the energy form used by both human and bacterial cells. The efficiency of the enzyme is critical and drugs can act that modulate the efficiency or inactivate it, thereby killing the cells. For this reason, the ATP synthase is a good target for drugs that could control obesity, diabetes, cancer, and act as antibiotics and the studies in this project will provide new avenues for drug discovery.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM066223-10
Application #
8542861
Study Section
Biochemistry and Biophysics of Membranes Study Section (BBM)
Program Officer
Anderson, Vernon
Project Start
2002-07-01
Project End
2016-06-30
Budget Start
2013-07-01
Budget End
2014-06-30
Support Year
10
Fiscal Year
2013
Total Cost
$286,285
Indirect Cost
$100,987
Name
Rosalind Franklin University
Department
Biochemistry
Type
Schools of Medicine
DUNS #
069501252
City
North Chicago
State
IL
Country
United States
Zip Code
60064
Steel, Bradley C; Nord, Ashley L; Wang, Yamin et al. (2015) Comparison between single-molecule and X-ray crystallography data on yeast F1-ATPase. Sci Rep 5:8773
Shah, Kalpit; Cheng, Yi; Hahn, Brian et al. (2015) Synonymous codon usage affects the expression of wild type and F508del CFTR. J Mol Biol 427:1464-79
Xu, Ting; Pagadala, Vijayakanth; Mueller, David M (2015) Understanding structure, function, and mutations in the mitochondrial ATP synthase. Microb Cell 2:105-125
Robinson, Graham C; Bason, John V; Montgomery, Martin G et al. (2013) The structure of F₁-ATPase from Saccharomyces cerevisiae inhibited by its regulatory protein IF₁. Open Biol 3:120164
Lieber, Daniel S; Calvo, Sarah E; Shanahan, Kristy et al. (2013) Targeted exome sequencing of suspected mitochondrial disorders. Neurology 80:1762-70
Bilyard, Thomas; Nakanishi-Matsui, Mayumi; Steel, Bradley C et al. (2013) High-resolution single-molecule characterization of the enzymatic states in Escherichia coli F1-ATPase. Philos Trans R Soc Lond B Biol Sci 368:20120023
Symersky, Jindrich; Osowski, Daniel; Walters, D Eric et al. (2012) Oligomycin frames a common drug-binding site in the ATP synthase. Proc Natl Acad Sci U S A 109:13961-5
Symersky, Jindrich; Pagadala, Vijayakanth; Osowski, Daniel et al. (2012) Structure of the c(10) ring of the yeast mitochondrial ATP synthase in the open conformation. Nat Struct Mol Biol 19:485-91, S1
Pagadala, Vijayakanth; Vistain, Luke; Symersky, Jindrich et al. (2011) Characterization of the mitochondrial ATP synthase from yeast Saccharomyces cerevisae. J Bioenerg Biomembr 43:333-47
Little, Max A; Steel, Bradley C; Bai, Fan et al. (2011) Steps and bumps: precision extraction of discrete states of molecular machines. Biophys J 101:477-85

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