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Subcellular localization of the enterobacterial common antigen GT‐E‐like glycosyltransferase, WecG

Enterobacterales have developed a specialized outer membrane polysaccharide (enterobacterial common antigen [ECA]). ECA biosynthesis begins on the cytoplasmic side of the inner membrane (IM) where glycosyltransferases sequentially add sugar moieties to form a complete repeat unit which is then trans...

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Autores principales: Maczuga, Nicholas, Tran, Elizabeth N. H., Morona, Renato
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9804384/
https://www.ncbi.nlm.nih.gov/pubmed/36006410
http://dx.doi.org/10.1111/mmi.14973
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author Maczuga, Nicholas
Tran, Elizabeth N. H.
Morona, Renato
author_facet Maczuga, Nicholas
Tran, Elizabeth N. H.
Morona, Renato
author_sort Maczuga, Nicholas
collection PubMed
description Enterobacterales have developed a specialized outer membrane polysaccharide (enterobacterial common antigen [ECA]). ECA biosynthesis begins on the cytoplasmic side of the inner membrane (IM) where glycosyltransferases sequentially add sugar moieties to form a complete repeat unit which is then translocated across the IM by WzxE before being polymerized into short linear chains by WzyE/WzzE. Research into WecG, the enzyme responsible for generating ECA lipid‐II, has not progressed beyond Barr et al. (1988) who described WecG as a membrane protein. Here we revise our understanding of WecG and re‐characterize it as a peripherally associated membrane protein. Through the use of Western immunoblotting we show that WecG in Shigella flexneri is maintained to the IM via its three C‐terminal helices and further identify key residues in helix II which are critical for this interaction which has allowed us to identify WecG as a GT‐E glycosyltransferase. We investigate the possibility of protein complexes and ultimately show that ECA lipid‐I maintains WecG to the membrane which is crucial for its function. This research is the first since Barr et al. (1988) to investigate the biochemistry of WecG and reveals possible novel drug targets to inhibit WecG and thus ECA function and cell viability.
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spelling pubmed-98043842023-01-03 Subcellular localization of the enterobacterial common antigen GT‐E‐like glycosyltransferase, WecG Maczuga, Nicholas Tran, Elizabeth N. H. Morona, Renato Mol Microbiol Research Articles Enterobacterales have developed a specialized outer membrane polysaccharide (enterobacterial common antigen [ECA]). ECA biosynthesis begins on the cytoplasmic side of the inner membrane (IM) where glycosyltransferases sequentially add sugar moieties to form a complete repeat unit which is then translocated across the IM by WzxE before being polymerized into short linear chains by WzyE/WzzE. Research into WecG, the enzyme responsible for generating ECA lipid‐II, has not progressed beyond Barr et al. (1988) who described WecG as a membrane protein. Here we revise our understanding of WecG and re‐characterize it as a peripherally associated membrane protein. Through the use of Western immunoblotting we show that WecG in Shigella flexneri is maintained to the IM via its three C‐terminal helices and further identify key residues in helix II which are critical for this interaction which has allowed us to identify WecG as a GT‐E glycosyltransferase. We investigate the possibility of protein complexes and ultimately show that ECA lipid‐I maintains WecG to the membrane which is crucial for its function. This research is the first since Barr et al. (1988) to investigate the biochemistry of WecG and reveals possible novel drug targets to inhibit WecG and thus ECA function and cell viability. John Wiley and Sons Inc. 2022-08-25 2022-10 /pmc/articles/PMC9804384/ /pubmed/36006410 http://dx.doi.org/10.1111/mmi.14973 Text en © 2022 The Authors. Molecular Microbiology published by John Wiley & Sons Ltd. https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc-nd/4.0/ (https://creativecommons.org/licenses/by-nc-nd/4.0/) License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non‐commercial and no modifications or adaptations are made.
spellingShingle Research Articles
Maczuga, Nicholas
Tran, Elizabeth N. H.
Morona, Renato
Subcellular localization of the enterobacterial common antigen GT‐E‐like glycosyltransferase, WecG
title Subcellular localization of the enterobacterial common antigen GT‐E‐like glycosyltransferase, WecG
title_full Subcellular localization of the enterobacterial common antigen GT‐E‐like glycosyltransferase, WecG
title_fullStr Subcellular localization of the enterobacterial common antigen GT‐E‐like glycosyltransferase, WecG
title_full_unstemmed Subcellular localization of the enterobacterial common antigen GT‐E‐like glycosyltransferase, WecG
title_short Subcellular localization of the enterobacterial common antigen GT‐E‐like glycosyltransferase, WecG
title_sort subcellular localization of the enterobacterial common antigen gt‐e‐like glycosyltransferase, wecg
topic Research Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9804384/
https://www.ncbi.nlm.nih.gov/pubmed/36006410
http://dx.doi.org/10.1111/mmi.14973
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