The subunit composition of the human NADH dehydrogenase obtained by rapid one-step immunopurification

James Murray, Bing Zhang, Steven W. Taylor, Devin Oglesbee, Eoin Fahy, Michael F. Marusich, Soumitra S. Ghosh, Roderick A. Capaldi

Research output: Contribution to journalArticle

86 Scopus citations

Abstract

Defects of the NADH dehydrogenase complex are predominantly manifested in mitochondrial diseases and are significantly associated with the development of many late onset neurological disorders such as Parkinson's disease. Here we describe an immunocapture procedure for isolating this multisubunit membrane-bound complex from human tissue. Using small amounts of immunoisolated protein, one-dimensional and two-dimensional gel electrophoresis, matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) peptide mass finger printing (PMF), and nanoflow liquid chromatography mass spectrometry/mass spectrometry (LC-MS/MS), we can resolve and identify the human homologues of 42 polypeptides detected so far in the more extensively studied beef heart complex I. These polypeptides include the GRIM-19 protein, which is claimed to be involved in apoptosis, a polypeptide first identified by gene screening as a neuronal protein, as well as a protein thought to be in differentiation linked processes. The concordance of data from human and bovine complex I isolated by different procedures adds to the certainty that these novel proteins of seemingly diverse function are a part of complex I.

Original languageEnglish (US)
Pages (from-to)13619-13622
Number of pages4
JournalJournal of Biological Chemistry
Volume278
Issue number16
DOIs
StatePublished - Apr 18 2003

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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    Murray, J., Zhang, B., Taylor, S. W., Oglesbee, D., Fahy, E., Marusich, M. F., Ghosh, S. S., & Capaldi, R. A. (2003). The subunit composition of the human NADH dehydrogenase obtained by rapid one-step immunopurification. Journal of Biological Chemistry, 278(16), 13619-13622. https://doi.org/10.1074/jbc.C300064200