title: Quantification of proteins using (13)C7-labeled and unlabeled iodoacetanilide by nano liquid chromatography/nanoelectrospray ionization and by selected reaction monitoring mass spectrometry.
abstract: The combination of cysteine-specific modifiers, iodoacetanilide (IAA) and (13)C7-labeled iodoacetanilide ((13)C7-IAA), has been applied to absolute quantification of proteins. The selected reaction monitoring (SRM) with the use of nano liquid chromatography/nanoelectrospray ionization ion trap mass spectrometry (nano LC/nano-ESI-IT-MS) analysis was applied to precise quantification of three commercial proteins. Good correlation was observed between the theoretical ratios and observed ratios for all these proteins both in a simple buffer solution and in a complex protein environment. Due to efficient tagging, this method does not require separate synthesis of isotope-labeled peptides for the SRM studies. Therefore, this method is expected to be a useful tool for proteomics research.
authors.affiliation: Laboratory of Molecular Signature Analysis, Division of Health Sciences, Osaka University Graduate School of Medicine, 1-7 Yamadaoka, Suita, Osaka 565-0871, Japan; Laboratory Chemicals Division, Wako Pure Chemical Industries, Ltd., 3-1-2 Doshomachi, Chuo-ku, Osaka, Osaka 540-8605, Japan.
authors.forename: Sadamu
authors.initials: S
authors.lastname: Kurono
authors.affiliation: Laboratory of Molecular Signature Analysis, Division of Health Sciences, Osaka University Graduate School of Medicine, 1-7 Yamadaoka, Suita, Osaka 565-0871, Japan; Laboratory Chemicals Division, Wako Pure Chemical Industries, Ltd., 3-1-2 Doshomachi, Chuo-ku, Osaka, Osaka 540-8605, Japan.
authors.forename: Yuka
authors.initials: Y
authors.lastname: Kaneko
authors.affiliation: 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: Shuji
authors.initials: S
authors.lastname: Matsuura
authors.affiliation: Department of Chemistry and Biochemistry, Texas Tech University, Lubbock, TX 79409-1061, USA; Department of Ophthalmology and Visual Sciences, Texas Tech University Health Sciences Center, Lubbock, TX 79430, USA. Electronic address: satomi.niwayama@ttu.edu.
authors.forename: Satomi
authors.initials: S
authors.lastname: Niwayama
mesh_terms: D000083:Acetanilides / Q000737:chemistry*; D000818:Animals; D002247:Carbon Isotopes / Q000737:chemistry; D002417:Cattle; D002853:Chromatography, Liquid / Q000379:methods; D007553:Isotope Labeling / Q000379:methods; D007768:Lactalbumin / Q000032:analysis; D010047:Ovalbumin / Q000032:analysis; D010455:Peptides / Q000032:analysis; D011506:Proteins / Q000032:analysis*; D040901:Proteomics / Q000379:methods; D012710:Serum Albumin, Bovine / Q000032:analysis; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods
chemical_list: D000083:Acetanilides; D002247:Carbon Isotopes; D010455:Peptides; D011506:Proteins; C545111:iodoacetanilide; D012710:Serum Albumin, Bovine; D010047:Ovalbumin; D007768:Lactalbumin
keywords: Cysteine modifiers; Proteomics; Quantitative analysis; Selected reaction monitoring (SRM); Stable isotope labeling