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Metabolic Labeling of Legionaminic Acid in Flagellin Glycosylation of Campylobacter jejuni Identifies Maf4 as a Putative Legionaminyl Transferase

Campylobacter jejuni is the major human food‐borne pathogen. Its bipolar flagella are heavily O‐glycosylated with microbial sialic acids and essential for its motility and pathogenicity. However, both the glycosylation of flagella and the exact contribution of legionaminic acid (Leg) to flagellar ac...

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Autores principales: Meng, Xianke, Boons, Geert‐Jan, Wösten, Marc M. S. M., Wennekes, Tom
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9298399/
https://www.ncbi.nlm.nih.gov/pubmed/34519150
http://dx.doi.org/10.1002/anie.202107181
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author Meng, Xianke
Boons, Geert‐Jan
Wösten, Marc M. S. M.
Wennekes, Tom
author_facet Meng, Xianke
Boons, Geert‐Jan
Wösten, Marc M. S. M.
Wennekes, Tom
author_sort Meng, Xianke
collection PubMed
description Campylobacter jejuni is the major human food‐borne pathogen. Its bipolar flagella are heavily O‐glycosylated with microbial sialic acids and essential for its motility and pathogenicity. However, both the glycosylation of flagella and the exact contribution of legionaminic acid (Leg) to flagellar activity is poorly understood. Herein, we report the development of a metabolic labeling method for Leg glycosylation on bacterial flagella with probes based on azide‐modified Leg precursors. The hereby azido‐Leg labeled flagellin could be detected by Western blot analysis and imaged on intact bacteria. Using the probes on C. jejuni and its isogenic maf4 mutant we also further substantiated the identification of Maf4 as a putative Leg glycosyltransferase. Further evidence was provided by UPLC–MS detection of labeled CMP‐Leg and an in silico model of Maf4. This method and the developed probes will facilitate the study of Leg glycosylation and the functional role of this modification in C. jejuni motility and invasiveness.
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spelling pubmed-92983992022-07-21 Metabolic Labeling of Legionaminic Acid in Flagellin Glycosylation of Campylobacter jejuni Identifies Maf4 as a Putative Legionaminyl Transferase Meng, Xianke Boons, Geert‐Jan Wösten, Marc M. S. M. Wennekes, Tom Angew Chem Int Ed Engl Communications Campylobacter jejuni is the major human food‐borne pathogen. Its bipolar flagella are heavily O‐glycosylated with microbial sialic acids and essential for its motility and pathogenicity. However, both the glycosylation of flagella and the exact contribution of legionaminic acid (Leg) to flagellar activity is poorly understood. Herein, we report the development of a metabolic labeling method for Leg glycosylation on bacterial flagella with probes based on azide‐modified Leg precursors. The hereby azido‐Leg labeled flagellin could be detected by Western blot analysis and imaged on intact bacteria. Using the probes on C. jejuni and its isogenic maf4 mutant we also further substantiated the identification of Maf4 as a putative Leg glycosyltransferase. Further evidence was provided by UPLC–MS detection of labeled CMP‐Leg and an in silico model of Maf4. This method and the developed probes will facilitate the study of Leg glycosylation and the functional role of this modification in C. jejuni motility and invasiveness. John Wiley and Sons Inc. 2021-10-27 2021-11-15 /pmc/articles/PMC9298399/ /pubmed/34519150 http://dx.doi.org/10.1002/anie.202107181 Text en © 2021 The Authors. Angewandte Chemie International Edition published by Wiley-VCH GmbH https://creativecommons.org/licenses/by/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Communications
Meng, Xianke
Boons, Geert‐Jan
Wösten, Marc M. S. M.
Wennekes, Tom
Metabolic Labeling of Legionaminic Acid in Flagellin Glycosylation of Campylobacter jejuni Identifies Maf4 as a Putative Legionaminyl Transferase
title Metabolic Labeling of Legionaminic Acid in Flagellin Glycosylation of Campylobacter jejuni Identifies Maf4 as a Putative Legionaminyl Transferase
title_full Metabolic Labeling of Legionaminic Acid in Flagellin Glycosylation of Campylobacter jejuni Identifies Maf4 as a Putative Legionaminyl Transferase
title_fullStr Metabolic Labeling of Legionaminic Acid in Flagellin Glycosylation of Campylobacter jejuni Identifies Maf4 as a Putative Legionaminyl Transferase
title_full_unstemmed Metabolic Labeling of Legionaminic Acid in Flagellin Glycosylation of Campylobacter jejuni Identifies Maf4 as a Putative Legionaminyl Transferase
title_short Metabolic Labeling of Legionaminic Acid in Flagellin Glycosylation of Campylobacter jejuni Identifies Maf4 as a Putative Legionaminyl Transferase
title_sort metabolic labeling of legionaminic acid in flagellin glycosylation of campylobacter jejuni identifies maf4 as a putative legionaminyl transferase
topic Communications
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9298399/
https://www.ncbi.nlm.nih.gov/pubmed/34519150
http://dx.doi.org/10.1002/anie.202107181
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