Profiling of strain R2866 with carbohydrate-based covalent probes

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...

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Bibliographic Details
Published inOrganic & biomolecular chemistry Vol. 19; no. 2; pp. 476 - 485
Main Authors Metier, Camille, Dow, Jennifer, Wootton, Hayley, Lynham, Steven, Wren, Brendan, Wagner, Gerd K
Format Journal Article
Published 21.01.2021
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Summary: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. Labelling of proteins in Haemophilus influenzae with covalent, carbohydrate-based probes produced distinctive target profiles in lysates vs . intact cells.
Bibliography:Electronic supplementary information (ESI) available: Synthetic protocols for fluorophore controls
additional Fig. S1-S6; LC-MS/MS analysis of SDS-page gel bands; spectroscopic data for compounds
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See DOI
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10.1039/d0ob01971b
ISSN:1477-0520
1477-0539
DOI:10.1039/d0ob01971b