The subunit composition of the human NADH dehydrogenase obtained by rapid one-step immunopurification.
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.
Murray
James
J
Department of Molecular Biology, University of Oregon, Eugene 97403, USA.
Zhang
Bing
B
Taylor
Steven W
SW
Oglesbee
Devin
D
Fahy
Eoin
E
Marusich
Michael F
MF
Ghosh
Soumitra S
SS
Capaldi
Roderick A
RA
D000818:Animals; D017209:Apoptosis; D002417:Cattle; D002853:Chromatography, Liquid; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D013058:Mass Spectrometry; D008928:Mitochondria / Q000378:metabolism; D009206:Myocardium / Q000378:metabolism; D009245:NADH Dehydrogenase / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism; D010455:Peptides / Q000737:chemistry; D011485:Protein Binding; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D018076:DNA, Complementary; D010455:Peptides; D009245:NADH Dehydrogenase