Profiling of: Haemophilus influenzae strain R2866 with carbohydrate-based covalent probes

Camille Metier; Jennifer Dow; Hayley Wootton; Steven Lynham; Brendan Wren; Gerd K. Wagner

doi:10.1039/d0ob01971b

Profiling of: Haemophilus influenzae strain R2866 with carbohydrate-based covalent probes

Camille Metier, Jennifer Dow, Hayley Wootton, Steven Lynham, Brendan Wren, Gerd K. Wagner^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

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. This journal is

Original language	English
Pages (from-to)	476-485
Number of pages	10
Journal	ORGANIC AND BIOMOLECULAR CHEMISTRY
Volume	19
Issue number	2
DOIs	https://doi.org/10.1039/d0ob01971b
Publication status	Published - 14 Jan 2021

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10.1039/d0ob01971b

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title = "Profiling of: Haemophilus influenzae strain R2866 with carbohydrate-based covalent probes",

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. This journal is ",

author = "Camille Metier and Jennifer Dow and Hayley Wootton and Steven Lynham and Brendan Wren and Wagner, {Gerd K.}",

note = "Funding Information: We thank King{\textquoteright}s College London for a studentship (to CM), the EPSRC National Mass Spectrometry Facility, Swansea, for the recording of mass spectra, and Dr Jon Cuccui (LSHTM) for helpful discussions. The LgtC-expressing clone NMC-41 was a gift from Prof Warren Wakarchuk (University of Alberta). Funding from the Biotechnology & Biological Sciences Research Council (grant BB/N001591/1, to BW) is gratefully acknowledged. Publisher Copyright: {\textcopyright} The Royal Society of Chemistry. Copyright: Copyright 2021 Elsevier B.V., All rights reserved.",

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doi = "10.1039/d0ob01971b",

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AU - Wren, Brendan

AU - Wagner, Gerd K.

N1 - Funding Information: We thank King’s College London for a studentship (to CM), the EPSRC National Mass Spectrometry Facility, Swansea, for the recording of mass spectra, and Dr Jon Cuccui (LSHTM) for helpful discussions. The LgtC-expressing clone NMC-41 was a gift from Prof Warren Wakarchuk (University of Alberta). Funding from the Biotechnology & Biological Sciences Research Council (grant BB/N001591/1, to BW) is gratefully acknowledged. Publisher Copyright: © The Royal Society of Chemistry. Copyright: Copyright 2021 Elsevier B.V., All rights reserved.

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Profiling of: Haemophilus influenzae strain R2866 with carbohydrate-based covalent probes

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