Method of extraction and proteome profiling of mycobacteria using liquid chromatography-high resolution mass spectrometry

摘要: Advances in massively parallel sequencing, of complete bacterial genomes, have led to many novel findings the field genomics. However, these data often lack correlation with expressed protein profiles. It has been demonstrated that even very closely related such as mycobacteria, express drastically different phenotypes. These phenotypes major roles pathogenicity. Therefore, it is just important a method for examining proteome bacterium well its genome. studies are further complicated mycobacteria due cell wall and mycolic acid. A comprehensive identification characterization whole mycobacterium profile needed. In present study, simple, sensitive, specific liquid chromatography tandem mass spectrometry was developed extraction, purification profiling mycobacterial various species. During development, sonication bead-beating lysis protocol tested using 15% Acetonitrile 6 M guanidine-HCl (GuHCl) extraction solvent. Sonication GuHCl glass beads preferred lysis. This reverse phase Q Exactive ™ Plus Orbitrap™ spectrometer peptide identification. Bottom-up chromatography-mass LC–MS analysis resulted greater than 2500 proteins.