1. [In vitro protein phosphorylation as a template for SRM method development].
- Author
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Zav'ialova MG, Zgoda VG, Kharybin ON, and Nikolaev EN
- Subjects
- Amino Acid Sequence, Animals, Brain Chemistry, Brain Neoplasms metabolism, Brain Neoplasms pathology, Cattle, Glioma metabolism, Glioma pathology, Humans, Mitogen-Activated Protein Kinase 1 metabolism, Molecular Sequence Data, Myelin Basic Protein metabolism, Peptide Mapping, Phosphopeptides metabolism, Phosphorylation, Swine, Brain Neoplasms chemistry, Glioma chemistry, Mitogen-Activated Protein Kinase 1 chemistry, Myelin Basic Protein chemistry, Phosphopeptides chemistry, Protein Processing, Post-Translational
- 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.
- Published
- 2014
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