In vitro protein phosphorylation as a template for SRM method development

   


1. Orekhovich Institute of Biomedical Chemistry (IBMC)
2. Orekhovich Institute of Biomedical Chemistry (IBMC); Institute for Energy Problems of Chemical Physics RAS, Moscow, Russia
Type: Experimental study
DOI: 10.18097/PBMC20146006668      UDK: 577.15.08      PubMed Id: 25552507
Year: 2014 vol: 60  issue:6  pages: 668-676
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.
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Reference: Zavialova M.G., Zgoda V.G., Kharybin O.N., Nikolayev E.N., In vitro protein phosphorylation as a template for SRM method development, Biomeditsinskaya khimiya, 2014, vol: 60(6), 668-676.
This paper is also available as the English translation:10.1134/S1990750815040095
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