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Proteomic Analysis of the Subunit Composition of Complex I (NADH:Ubiquinone Oxidoreductase) From Bovine Heart Mitochondria

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Cardiovascular Proteomics

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 357))

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Abstract

Complex I from the inner membranes of mammalian mitochondria is a complicated membrane-bound assembly of redox centers (flavin mononucleotide cofactor, iron sulphur centers) andat least 46 different proteins. The hydrophobic nature of its membranebound subunits and the complexity of subunit content present a substantial analytical challenge. The complete protein chemical analysis of complex I purified from bovine mitochondria required the resolution of subunits by one-dimensional and two-dimensional electrophoresis, reverse-phase chromatography, and combinations of these techniques. These subunits were characterized by mass spectrometry (MS)-based protein identification methods, requiring both peptide mass fingerprinting and amino acid sequencing by tandem MS. The components were identified also and characterized by measurements of subunit molecular mass. These strategies have provided a comprehensive view of the subunit content of the intact complex, its structural domains, and stable subunit modifications.

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Author information

Authors and Affiliations

  1. Dunn Human Nutrition Unit, The Medical Research Council, Cambridge, UK

    Ian M. Fearnley, Joe Carroll & John E. Walker

Authors
  1. Ian M. Fearnley
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  2. Joe Carroll
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  3. John E. Walker
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Editor information

Editors and Affiliations

  1. Department of Immunology, Fundación Jiménez Díaz, Universidad Complutense, Madrid, Spain

    Fernando Vivanco

  2. Department of Biochemistry and Molecular Biology I, Universidad Complutense, Madrid, Spain

    Fernando Vivanco

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© 2007 Humana Press Inc.

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Fearnley, I.M., Carroll, J., Walker, J.E. (2007). Proteomic Analysis of the Subunit Composition of Complex I (NADH:Ubiquinone Oxidoreductase) From Bovine Heart Mitochondria. In: Vivanco, F. (eds) Cardiovascular Proteomics. Methods in Molecular Biology™, vol 357. Humana Press. https://doi.org/10.1385/1-59745-214-9:103

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  • DOI: https://doi.org/10.1385/1-59745-214-9:103

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-535-4

  • Online ISBN: 978-1-59745-214-4

  • eBook Packages: Springer Protocols

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