Mass spectrometric identification of a novel phosphorylation site in subunit NDUFA10 of bovine mitochondrial complex I

• Published in: FEBS letters 2005-04, Vol.579 (11), p.2485-2490
• Main Authors: Schilling, Birgit, Aggeler, Robert, Schulenberg, Birte, Murray, James, Row, Richard H., Capaldi, Roderick A., Gibson, Bradford W.
• Format: Article
• Language: English
• Subjects: Alkaline Phosphatase - metabolism; Amino Acid Sequence; Animals; Cattle; Complex I; Electron Transport Complex I - chemistry; Electron Transport Complex I - metabolism; Humans; Mass spectrometry; Mitochondria; Molecular Sequence Data; Parkinson's disease; Phosphorylation; Protein Subunits - chemistry; Protein Subunits - metabolism; Sequence Alignment; Spectrometry, Mass, Electrospray Ionization
• ISSN: 0014-5793; 1873-3468
• DOI: 10.1016/j.febslet.2005.03.061

Mitochondrial Complex I (NADH:ubiquinone oxidoreductase) consists of at least 46 subunits. Phosphorylation of the 42-kDa subunit NDUFA10 was recently reported using a novel phosphoprotein stain [Schulenberg et al. (2003) Analysis of steady-state protein phosphorylation in mitochondria using a novel fluorescent phosphosensor dye. J. Biol. Chem. 278, 27251]. Two smaller Complex I phosphoproteins, ESSS and MWFE, and their sites of modification, have since been determined [Chen et al. (2004) The phosphorylation of subunits of complex I from bovine heart mitochondria. J. Biol. Chem. 279, 26036]. Here we identify the site of phosphorylation in NDUFA10 from bovine heart mitochondria by tandem mass spectrometry. A single phosphopeptide spanning residues 47–60 was identified and confirmed by synthesis to be (47)LITVDGNICSGKpSK(60), establishing serine-59 as the site of phosphorylation.