Analysis of flagellar proteins in Clostridium difficile isolates of clinically relevant PCR-ribotypes

Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Kateřina Houdková MPharm Supervisor: RNDr. Klára Konečná, Ph.D. Consultant: PharmDr. Jiří Dresler, Ph.D. Title of rigorous thesis: Analysis of flagellar proteins in C. difficile isolates of clinically relevant PCR-ribotypes Background: Strains of C. difficile of known human epidemiologic importance are associated with severe clinical features of C. difficile infection (CDI). In this study, a panel of eight different PCR-ribotypes (RTs) with their proteins released in vitro were subjected to analysis. The aim of this work is to monitor the relationship between secretions of individual proteins associated with flagellar formation and function in C. difficile strains of variable virulence. Methods: Within our research, a combination of tandem mass spectrometry with liquid chromatography was used. The semi-quantitative analysis employed label free quantification (LFQ) approach. Results: From the quantifiable proteins, 17 were significantly increased in functional annotations. Among them, several known factors connected with flagellar assembly and other functions were identified. Higher expression of selected flagellar proteins clearly distinguished RTs 027, 176, 005 and 012, confirming the pathogenic..