Profiling of Haemophilus influenzae strain R2866 with carbohydrate-based covalent probes. Issue 2 (23rd December 2020)

• Record Type: Journal Article
• Authors: Metier, Camille; Dow, Jennifer; Wootton, Hayley; Lynham, Steven; Wren, Brendan; Wagner, Gerd K.
• Is Part Of: Organic & biomolecular chemistry. Volume 19:Issue 2(2020)
• Page Start: 476
• Page End: 485
• Publication Date: 2020-12-23
• Subjects: Chemistry, Organic -- Periodicals; Bioorganic chemistry -- Periodicals; Chemistry, Physical organic -- Periodicals; 547
• DOI: 10.1039/d0ob01971b
• Languages: English
• ISSNs: 1477-0520

Abstract

Abstract : Labelling of proteins in Haemophilus influenzae with covalent, carbohydrate-based probes produced distinctive target profiles in lysates vs . intact cells. Abstract : We demonstrate the application of four covalent probes based on anomerically pure d -galactosamine and d -glucosamine scaffolds for the profiling of Haemophilus influenzae strain R2866. The probes have been used successfully for the labelling of target proteins not only in cell lysates, but also in intact cells. Differences in the labelling patterns between lysates and intact cells indicate that the probes can penetrate into the periplasm, but not the cytoplasm of H. influenzae . Analysis of selected target proteins by LC-MS/MS suggests predominant labelling of nucleotide-binding proteins, including several known antibacterial drug targets. Our protocols will aid the identification of molecular determinants of bacterial pathogenicity in Haemophilus influenzae and other bacterial pathogens.