The long-range goal of this project is to understand the functional role and unusual association of phospholipids at individual binding sites on mitochondrial membrane proteins. Towards this aim, emphasis will be placed on determining the association of a unique mitochondrial phospholipid, cardiolipin, with a number of proteins known to have a functional dependence on cardiolipin for their biological function. Three types of mitochondrial membrane associated proteins will be studied: 1) the multisubunit electron transport complexes (cytochrome c oxidase and cytochrome bc1); 2) the dimeric inner membrane translocases (ADP/ATP translocase); and 3) the presequences of two cytoplasmically synthesized and mitochondrially targeted proteins (the synthetic presequences of yeast cytochrome c oxidase subunit IV and human ornithine transcarbamylase). The approach to be taken is to directly study the structural features of the cardiolipin binding site within each of these purified proteins using a series of novel synthetic cardiolipin derivatives. The synthetic cardiolipin analogues will contain chemically reactive groups, photoreactive groups (arylazides), fluorescent labels (dansyl and fluorescein) and easily detected groups (biotin). Several derivatives will be di-substituted so that cardiolipin can be used to covalently label subunits with biotin and fluorescent probes, and to crosslink subunits and sequences interacting with its apolar and polar regions. Some of these cardiolipin derivatives have been prepared during the last period of funding, others will be synthesized during the next phase of the project. Understanding the unique association of cardiolipin with one of these proteins, cytochrome c oxidase, has been one of the major objectives of our research during the past two periods of funding. The current proposal extends these studies to include other mitochondrial enzymes and proteins that are functionally dependent upon cardiolipin. By comparing the association of synthetic cardiolipin analogues with several different membrane proteins, structural features that are common among these cardiolipin dependent proteins will be determined, and the molecular basis of cardiolipin participation in mitochondrial function will be established

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM024795-16
Application #
3272523
Study Section
Biophysical Chemistry Study Section (BBCB)
Project Start
1978-01-01
Project End
1994-12-31
Budget Start
1993-01-01
Budget End
1993-12-31
Support Year
16
Fiscal Year
1993
Total Cost
Indirect Cost
Name
University of Texas Health Science Center San Antonio
Department
Type
Schools of Dentistry
DUNS #
800772162
City
San Antonio
State
TX
Country
United States
Zip Code
78229
Musatov, Andrey; Siposova, Katarina; Kubovcikova, Martina et al. (2016) Functional and structural evaluation of bovine heart cytochrome c oxidase incorporated into bicelles. Biochimie 121:21-8
Musatov, Andrej; Varha?, Rastislav; Hosler, Jonathan P et al. (2016) Delipidation of cytochrome c oxidase from Rhodobacter sphaeroides destabilizes its quaternary structure. Biochimie 125:23-31
Musatov, Andrej; Robinson, Neal C (2014) Bound cardiolipin is essential for cytochrome c oxidase proton translocation. Biochimie 105:159-64
Musatov, Andrej; Fabian, Marian; Varha?, Rastislav (2013) Elucidating the mechanism of ferrocytochrome c heme disruption by peroxidized cardiolipin. J Biol Inorg Chem 18:137-44
Musatov, Andrej (2013) Dual effect of heparin on Fe²?-induced cardiolipin peroxidation: implications for peroxidation of cytochrome c oxidase bound cardiolipin. J Biol Inorg Chem 18:729-37
Musatov, Andrej; Robinson, Neal C (2012) Susceptibility of mitochondrial electron-transport complexes to oxidative damage. Focus on cytochrome c oxidase. Free Radic Res 46:1313-26
Sedlak, Erik; Fabian, Marian; Robinson, Neal C et al. (2010) Ferricytochrome c protects mitochondrial cytochrome c oxidase against hydrogen peroxide-induced oxidative damage. Free Radic Biol Med 49:1574-81
Varhac, Rastislav; Robinson, Neal C; Musatov, Andrej (2009) Removal of bound Triton X-100 from purified bovine heart cytochrome bc1. Anal Biochem 395:268-70
Sedlák, Erik; Robinson, Neal C (2009) Sequential dissociation of subunits from bovine heart cytochrome C oxidase by urea. Biochemistry 48:8143-50
Lemma-Gray, Patrizia; Valusova, Eva; Carroll, Christopher A et al. (2008) Subunit analysis of bovine heart complex I by reversed-phase high-performance liquid chromatography, electrospray ionization-tandem mass spectrometry, and matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry. Anal Biochem 382:116-21

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