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Study of proteomic composition of exhaled breath condensate (EBC) is a promising non-invasive method for diagnostics of respiratory system diseases in patients. In this study the EBC proteomic composition of 53 donors, including patients with different respiratory system diseases has been investigated. Cytoskeletal keratins type II (1, 2, 3, 4, 5, 6) and cytoskeletal keratins type I (9, 10, 14, 15, 16) were invariant for all samples. Analyzing the frequency of occurrence of proteins in different groups of examined patients, several categories of proteins have been recognized: proteins found in all pathologies (dermcidin, alpha-1- microglobulin, SHROOM3), proteins simultaneously found in two groups (CSTA, LCN1, JUP, PIP, TXN), and proteins specific for a particular group (PRDX1, annexin A1/A2). The EBC analysis by HPLC-MS/MS can be used for identification of potential protein markers specific for inflammatory pulmonary diseases of infective origin (pneumonia) as well as for non-infectious diseases such as chronic obstructive pulmonary disease (COPD).

Phosphorylation is one of the most common and the most important posttranslational modifications (PTM) of proteins. The study of protein phosphoforms is complicated by their low abundance in analyzed biological samples compared with the abundance of corresponding unmodified proteins. The method of selected reactions monitoring (SRM) allows to perform very sensitive and selective PTM analysis with high specificity and sensitivity. Using myelin basic protein (MBP) as a model we have developed a method for phosphoprotein detection by SRM. The method is based on obtaining phosphoproteins in a reconstituted kinase system and following usage of tryptic fragments of the phosphorylated protein as a template for the development of the SRM method. The developed method has been successfully applied for detection of phosphopeptides of myelin basic protein in human brain glioma samples.