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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| Published in | Organic & biomolecular chemistry Vol. 19; no. 2; pp. 476 - 485 |
|---|---|
| Main Authors | , , , , , |
| Format | Journal Article |
| Published |
21.01.2021
|
| Online Access | Get full text |
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| 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.
Labelling of proteins in
Haemophilus influenzae
with covalent, carbohydrate-based probes produced distinctive target profiles in lysates
vs
. intact cells. |
|---|---|
| AbstractList | 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. |
| Author | Wagner, Gerd K Dow, Jennifer Metier, Camille Wootton, Hayley Lynham, Steven Wren, Brendan |
| AuthorAffiliation | Department of Chemistry Queen's University Belfast Centre of Excellence for Mass Spectrometry The James Black Centre King's College London London School of Hygiene and Tropical Medicine School of Pharmacy Proteomics Facility Department of Infection Biology Medical Biology Centre |
| AuthorAffiliation_xml | – name: Department of Infection Biology – name: Department of Chemistry – name: Centre of Excellence for Mass Spectrometry – name: Queen's University Belfast – name: School of Pharmacy – name: King's College London – name: Medical Biology Centre – name: The James Black Centre – name: London School of Hygiene and Tropical Medicine – name: Proteomics Facility |
| Author_xml | – sequence: 1 givenname: Camille surname: Metier fullname: Metier, Camille – sequence: 2 givenname: Jennifer surname: Dow fullname: Dow, Jennifer – sequence: 3 givenname: Hayley surname: Wootton fullname: Wootton, Hayley – sequence: 4 givenname: Steven surname: Lynham fullname: Lynham, Steven – sequence: 5 givenname: Brendan surname: Wren fullname: Wren, Brendan – sequence: 6 givenname: Gerd K surname: Wagner fullname: Wagner, Gerd K |
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| ContentType | Journal Article |
| DOI | 10.1039/d0ob01971b |
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| Notes | 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 and 9 2-7 , See DOI 10 10.1039/d0ob01971b |
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| References_xml | – publication-title: Profiling protein expression in Klebsiella pneumoniae with a carbohydrate-based probe doi: Metier Peng Xu Wootton Riesi Lynham Zadi Turner Wand Sutton Wagner – issn: 2019 issue: 420 end-page: 23-48 publication-title: Current Topics in Microbiology and Immunology doi: Sharifzadeh Shirley Carlson – issn: 2017 issue: 1491 end-page: 57-74 publication-title: Methods in Molecular Biology doi: Krysiak Sieber |
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d
-galactosamine and
d
-glucosamine scaffolds for the profiling of... |
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| Title | Profiling of strain R2866 with carbohydrate-based covalent probes |
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