Definition of the mitochondrial proteome by measurement of molecular masses of membrane proteins.

TitleDefinition of the mitochondrial proteome by measurement of molecular masses of membrane proteins.
Publication TypeJournal Article
Year of Publication2006
AuthorsCarroll, J, Fearnley, IM, Walker, JE
JournalProc Natl Acad Sci U S A
Volume103
Issue44
Pagination16170-5
Date Published2006 Oct 31
ISSN0027-8424
KeywordsAnimals, Cattle, Chromatography, Liquid, Hydrophobic and Hydrophilic Interactions, Mass Spectrometry, Membrane Proteins, Mitochondria, Heart, Protein Subunits, Proteome, Solvents, Spectrometry, Mass, Electrospray Ionization
Abstract

The covalent structure of a protein is incompletely defined by its gene sequence, and mass spectrometric analysis of the intact protein is needed to detect the presence of any posttranslational modifications. Because most membrane proteins are purified in detergents that are incompatible with mass spectrometric ionization techniques, this essential measurement has not been made on many hydrophobic proteins, and so proteomic data are incomplete. We have extracted membrane proteins from bovine mitochondria and detergent-purified NADH:ubiquinone oxidoreductase (complex I) with organic solvents, fractionated the mixtures by hydrophilic interaction chromatography, and measured the molecular masses of the intact membrane proteins, including those of six subunits of complex I that are encoded in mitochondrial DNA. These measurements resolve long-standing uncertainties about the interpretation of the mitochondrial genome, and they contribute significantly to the definition of the covalent composition of complex I.

DOI10.1073/pnas.0607719103
Alternate JournalProc. Natl. Acad. Sci. U.S.A.
Citation Key10.1073/pnas.0607719103
PubMed ID17060615
PubMed Central IDPMC1621045
Grant ListMC_U105663148 / / Medical Research Council / United Kingdom