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Published online
doi:10.1083/jcb.200801152
The Journal of Cell Biology, Vol. 182, No. 5, 937-950
The Rockefeller University Press, 0021-9525 $30.00
© Claypool et al.
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Article

Cardiolipin defines the interactome of the major ADP/ATP carrier protein of the mitochondrial inner membrane



Steven M. Claypool1, Yavuz Oktay1, Pinmanee Boontheung1, Joseph A. Loo1,2,3, and Carla M. Koehler1,3

1 Department of Chemistry and Biochemistry, 2 Department of Biological Chemistry, David Geffen School of Medicine, and 3 the Molecular Biology Institute, University of California, Los Angeles, Los Angeles, CA 90095

Correspondence to Carla M. Koehler: Koehler{at}chem.ucla.edu

Defined mutations in the mitochondrial ADP/ATP carrier (AAC) are associated with certain types of progressive external ophthalmoplegia. AAC is required for oxidative phosphorylation (OXPHOS), and dysregulation of AAC has been implicated in apoptosis. Little is known about the AAC interactome, aside from a known requirement for the phospholipid cardiolipin (CL) and that it is thought to function as a homodimer. Using a newly developed dual affinity tag, we demonstrate that yeast AAC2 physically participates in several protein complexes of distinct size and composition. The respiratory supercomplex and several smaller AAC2-containing complexes, including other members of the mitochondrial carrier family, are identified here. In the absence of CL, most of the defined interactions are destabilized or undetectable. The absence of CL and/or AAC2 results in distinct yet additive alterations in respiratory supercomplex structure and respiratory function. Thus, a single lipid can significantly alter the functional interactome of an individual protein.

Abbreviations used in this paper: AAC, ADP/ATP carrier; BN-PAGE, blue native polyacrylamide gel electrophoresis; CL, cardiolipin; CNAP, consecutive nondenaturing affinity purification; cyt c, cytochrome c; ETC, electron transport chain; IM, inner membrane; IP, immunoprecipitate; LC–MS/MS, liquid chromatography–tandem mass spectrometry; OXPHOS, oxidative phosphorylation; PEO, progressive external ophthalmoplegia; RCR, respiratory control ratio; Taz1p, tafazzin; wt, wild type.

© 2008 Claypool et al. This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.jcb.org/misc/terms.shtml). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).


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