Abstract
We have mapped the sites of ischemia/reperfusion-induced phosphorylation of cytochrome c oxidase (CcO) subunits in rabbit hearts by using a combination of Blue Native gel/Tricine gel electrophoresis and nano-LC-MS/MS approaches. We used precursor ion scanning combined with neutral loss scanning and found that mature CcO subunit I was phosphorylated at tandem Ser115/Ser116 positions, subunit IVi1 at Thr52 and subunit Vb at Ser40. These sites are highly conserved in mammalian species. Molecular modeling suggests that phosphorylation sites of subunit I face the inter membrane space while those of subunits IVi1 and Vb face the matrix side.
Publication types
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Research Support, N.I.H., Extramural
MeSH terms
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Amino Acid Sequence
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Animals
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Chromatography, Liquid
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Electron Transport Complex IV / chemistry*
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Electron Transport Complex IV / metabolism
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Electrophoresis, Polyacrylamide Gel
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Glycine / analogs & derivatives
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Glycine / chemistry
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Heart
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Molecular Sequence Data
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Myocardial Reperfusion Injury / enzymology*
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Myocardium / enzymology*
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Phosphorylation
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Protein Conformation
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Protein Structure, Tertiary
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Protein Subunits / chemistry
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Protein Subunits / metabolism
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Rabbits
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Serine / metabolism
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Spectrometry, Mass, Electrospray Ionization
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Threonine / metabolism
Substances
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Protein Subunits
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Threonine
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Serine
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Electron Transport Complex IV
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Glycine
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tricine