title: [In vitro protein phosphorylation as a template for SRM method development].
abstract: Phosphorylation is one of the most common posttranslational modification (PTM) of proteins. Main challenge of phosphoprotein detection is their low abundance comparing to abundance of unmodified proteins. The method of selected reactions monitoring (SRM) allows to perform very sensitive and selective analysis of desired PTMs. Using myelin basic protein (MBP) as a model we have developed a method for phosphoprotein detection by SRM. The method is based on obtaining of phosphoproteins in a reconstituted kinase system and following usage these phosphorylated protein as a template for the development of the SRM method. The developed method was successfully applied for detection of phosphopeptides of myelin basic protein in the samples of human brain glioma.
authors.affiliation: Orekhovich Institute of Biomedical Chemistry (IBMC).
authors.forename: M G
authors.initials: MG
authors.lastname: Zav'ialova
authors.affiliation: Orekhovich Institute of Biomedical Chemistry (IBMC).
authors.forename: V G
authors.initials: VG
authors.lastname: Zgoda
authors.affiliation: Orekhovich Institute of Biomedical Chemistry (IBMC).
authors.forename: O N
authors.initials: ON
authors.lastname: Kharybin
authors.affiliation: Orekhovich Institute of Biomedical Chemistry (IBMC); Institute for Energy Problems of Chemical Physics RAS, Moscow, Russia.
authors.forename: E N
authors.initials: EN
authors.lastname: Nikolaev
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001923:Brain Chemistry; D001932:Brain Neoplasms / Q000737:chemistry* / Q000378:metabolism / Q000473:pathology; D002417:Cattle; D005910:Glioma / Q000737:chemistry* / Q000378:metabolism / Q000473:pathology; D006801:Humans; D019950:Mitogen-Activated Protein Kinase 1 / Q000737:chemistry* / Q000378:metabolism; D008969:Molecular Sequence Data; D004676:Myelin Basic Protein / Q000737:chemistry* / Q000378:metabolism; D010449:Peptide Mapping; D010748:Phosphopeptides / Q000737:chemistry* / Q000378:metabolism; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational; D013552:Swine
chemical_list: D004676:Myelin Basic Protein; D010748:Phosphopeptides; C535150:MAPK1 protein, human; D019950:Mitogen-Activated Protein Kinase 1
keywords: post-translational modifications (PTM); protein phosphorylation; proteomics; selected reaction monitoring (SRM)