The mitochondrial F1Fo ATP synthase is responsible for the synthesis 90% of the ATP under aerobic conditions. The mitochondrial 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. Despite many peopledecades devoted across the globe, the structure of the entire enzyme complex has remained elusive. Because of recent technological advances and progress, we are now on the brink of obtaining the high-resolution structure of the mitochondrial enzyme. One key unanswered question is: what are the molecular determinants that cause the central stalk to rotate during ATP synthesis and hydrolysis? A principal goal of this project is to determine the high-resolution crystal structure of the eukaryotic ATP synthase complex.
This aim will give critical structural details into the understanding of the mechanism of proton translocation coupled to rotation driving ATP synthesis or driven by ATP hydrolysis. We will investigate the structural relationship of the ATP synthase in a number of reaction intermediate structures with inhibitors bound. We will exploit our understanding of the structure and biochemistry of the ATP synthase and aim to gain a further understanding of human mitochondrial diseases and variations amongst the population. This study will help bridge the knowledge gap between the genomic studies and biochemistry and will allow for translation into healthcare. Overall, the project will provide basic understanding on the mechanism of proton translocation coupled to an energy-requiring event ? the synthesis of ATP from ADP and Pi ? and aid in knowledge-based decisions in both the health and disease.

Public Health Relevance

ThisstudywilladvanceourunderstandingofthemechanismofATPsynthaseand couplingofiontranslocationtothesynthesisofATP.Coupledwithbiochemical studiesontheeffectofdiseasecausingmutationsthataffecttheenzymefunction, theresultshereinwillleadtoadvancesinthetreatmentofdiseasesandadverse healthconditions.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
3R01GM066223-15S1
Application #
9698656
Study Section
Macromolecular Structure and Function C Study Section (MSFC)
Program Officer
Anderson, Vernon
Project Start
2002-07-01
Project End
2019-06-30
Budget Start
2018-07-01
Budget End
2019-06-30
Support Year
15
Fiscal Year
2018
Total Cost
Indirect Cost
Name
Rosalind Franklin University
Department
Biochemistry
Type
Schools of Medicine
DUNS #
069501252
City
North Chicago
State
IL
Country
United States
Zip Code
60064
Srivastava, Anurag P; Luo, Min; Zhou, Wenchang et al. (2018) High-resolution cryo-EM analysis of the yeast ATP synthase in a lipid membrane. Science 360:
Xu, Ting; Pagadala, Vijayakanth; Mueller, David M (2015) Understanding structure, function, and mutations in the mitochondrial ATP synthase. Microb Cell 2:105-125
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 427:1464-1479
Lieber, Daniel S; Calvo, Sarah E; Shanahan, Kristy et al. (2013) Targeted exome sequencing of suspected mitochondrial disorders. Neurology 80:1762-70
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
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

Showing the most recent 10 out of 21 publications