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Characterization of a Coenzyme Q biosynthetic complex in Saccharomyces cerevisiae
Author(s) -
Allan Christopher Michael,
Tran UyenPhuong,
Gulmezian Melissa,
Marbois Beth,
Clarke Catherine
Publication year - 2010
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.24.1_supplement.849.1
Subject(s) - coenzyme q – cytochrome c reductase , biochemistry , saccharomyces cerevisiae , yeast , inner mitochondrial membrane , immunoprecipitation , membrane protein , biology , integral membrane protein , mitochondrial carrier , mitochondrion , cofactor , mitochondrial matrix , molecular mass , chemistry , bacterial outer membrane , enzyme , membrane , gene , cytochrome c , escherichia coli , cytosol
Coenzyme Q (Q) is a critical molecule for proper functioning of the electron transport chain in mitochondrial respiration. Q is synthesized by a series of enzymes in the mitochondria and the human Q biosynthetic proteins are well conserved in the yeast Saccharomyces cerevisiae , with several capable of functionally substituting for their yeast homologues. Yeast is thus a good model for understanding Q biosynthesis in higher eukaryotes. There is evidence that several of the Q biosynthetic proteins, the Coq proteins, are associated in a large complex in the mitochondria. All of the Coq proteins are peripheral membrane proteins on the matrix face of the inner mitochondrial membrane, except for Coq2p which is an integral membrane protein. It is thus possible that Coq2p serves as the membrane anchor for the other Coq polypeptides in the complex. Coq2p has also been demonstrated to exist in a high molecular mass complex via gel filtration chromatography and 2D Blue Native‐PAGE, although its partner proteins have not been identified. One of our aims is to characterize the Coq2p‐containing complex to determine its role in the organization and stabilization of the complex. BN‐PAGE will be used to determine whether other Coq proteins co‐migrate with Coq2p in a high molecular mass complex. Immunoprecipitation of Coq2p tagged with the HA epitope will identify interacting proteins.