title: Quantitative protein analysis using (13)C7-labeled iodoacetanilide and d5-labeled N-ethylmaleimide by nano liquid chromatography/nanoelectrospray ionization ion trap mass spectrometry.
abstract: We have developed a methodology for quantitative analysis and concurrent identification of proteins by the modification of cysteine residues with a combination of iodoacetanilide (IAA, 1) and (13)C7-labeled iodoacetanilide ((13)C7-IAA, 2), or N-ethylmaleimide (NEM, 3) and d5-labeled N-ethylmaleimide (d5-NEM, 4), followed by mass spectrometric analysis using nano liquid chromatography/nanoelectrospray ionization ion trap mass spectrometry (nano LC/nano-ESI-IT-MS). The combinations of these stable isotope-labeled and unlabeled modifiers coupled with LC separation and ESI mass spectrometric analysis allow accurate quantitative analysis and identification of proteins, and therefore are expected to be a useful tool for proteomics research.
authors.affiliation: Joint Research Laboratory of Molecular Signature Analysis, Division of Health Sciences, Osaka University Graduate School of Medicine, 1-7 Yamadaoka, Suita, Osaka 565-0871, Japan.
authors.forename: Sadamu
authors.initials: S
authors.lastname: Kurono
authors.forename: Yuka
authors.initials: Y
authors.lastname: Kaneko
authors.forename: Satomi
authors.initials: S
authors.lastname: Niwayama
mesh_terms: D000083:Acetanilides / Q000737:chemistry*; D000818:Animals; D002247:Carbon Isotopes; D002417:Cattle; D002853:Chromatography, Liquid; D005033:Ethylmaleimide / Q000737:chemistry*; D007768:Lactalbumin / Q000032:analysis*; D015394:Molecular Structure; D036103:Nanotechnology; D010047:Ovalbumin / Q000032:analysis*; D012710:Serum Albumin, Bovine / Q000032:analysis*; D021241:Spectrometry, Mass, Electrospray Ionization
chemical_list: D000083:Acetanilides; D002247:Carbon Isotopes; C545111:iodoacetanilide; D012710:Serum Albumin, Bovine; D010047:Ovalbumin; D007768:Lactalbumin; D005033:Ethylmaleimide