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)
Type
Research Project (R01)
Project #
5R01GM066223-11
Application #
8685994
Study Section
Biochemistry and Biophysics of Membranes Study Section (BBM)
Program Officer
Anderson, Vernon
Project Start
Project End
Budget Start
Budget End
Support Year
11
Fiscal Year
2014
Total Cost
Indirect Cost
Name
Rosalind Franklin University of Medicine & Sci
Department
Biochemistry
Type
Schools of Medicine
DUNS #
City
North Chicago
State
IL
Country
United States
Zip Code
60064
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
Robinson, Graham C; Bason, John V; Montgomery, Martin G et al. (2013) The structure of F?-ATPase from Saccharomyces cerevisiae inhibited by its regulatory protein IF?. Open Biol 3:120164
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
Arsenieva, Diana; Symersky, Jindrich; Wang, Yamin et al. (2010) Crystal structures of mutant forms of the yeast F1 ATPase reveal two modes of uncoupling. J Biol Chem 285:36561-9
Notari, Luigi; Arakaki, Naokatu; Mueller, David et al. (2010) Pigment epithelium-derived factor binds to cell-surface F(1)-ATP synthase. FEBS J 277:2192-205
Sowa, Yoshiyuki; Steel, Bradley C; Berry, Richard M (2010) A simple backscattering microscope for fast tracking of biological molecules. Rev Sci Instrum 81:113704
Kabaleeswaran, Venkataraman; Shen, Hong; Symersky, Jindrich et al. (2009) Asymmetric structure of the yeast F1 ATPase in the absence of bound nucleotides. J Biol Chem 284:10546-51

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