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Molecular basis for polysaccharide recognition and modulated ATP hydrolysis by the O antigen ABC transporter
O antigens are ubiquitous protective extensions of lipopolysaccharides in the extracellular leaflet of the Gram-negative outer membrane. Following biosynthesis in the cytosol, the lipid-linked polysaccharide is transported to the periplasm by the WzmWzt ABC transporter. Often, O antigen secretion re...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9445017/ https://www.ncbi.nlm.nih.gov/pubmed/36064941 http://dx.doi.org/10.1038/s41467-022-32597-2 |
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author | Spellmon, Nicholas Muszyński, Artur Górniak, Ireneusz Vlach, Jiri Hahn, David Azadi, Parastoo Zimmer, Jochen |
author_facet | Spellmon, Nicholas Muszyński, Artur Górniak, Ireneusz Vlach, Jiri Hahn, David Azadi, Parastoo Zimmer, Jochen |
author_sort | Spellmon, Nicholas |
collection | PubMed |
description | O antigens are ubiquitous protective extensions of lipopolysaccharides in the extracellular leaflet of the Gram-negative outer membrane. Following biosynthesis in the cytosol, the lipid-linked polysaccharide is transported to the periplasm by the WzmWzt ABC transporter. Often, O antigen secretion requires the chemical modification of its elongating terminus, which the transporter recognizes via a carbohydrate-binding domain (CBD). Here, using components from A. aeolicus, we identify the O antigen structure with methylated mannose or rhamnose as its cap. Crystal and cryo electron microscopy structures reveal how WzmWzt recognizes this cap between its carbohydrate and nucleotide-binding domains in a nucleotide-free state. ATP binding induces drastic conformational changes of its CBD, terminating interactions with the O antigen. ATPase assays and site directed mutagenesis reveal reduced hydrolytic activity upon O antigen binding, likely to facilitate polymer loading into the ABC transporter. Our results elucidate critical steps in the recognition and translocation of polysaccharides by ABC transporters. |
format | Online Article Text |
id | pubmed-9445017 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-94450172022-09-07 Molecular basis for polysaccharide recognition and modulated ATP hydrolysis by the O antigen ABC transporter Spellmon, Nicholas Muszyński, Artur Górniak, Ireneusz Vlach, Jiri Hahn, David Azadi, Parastoo Zimmer, Jochen Nat Commun Article O antigens are ubiquitous protective extensions of lipopolysaccharides in the extracellular leaflet of the Gram-negative outer membrane. Following biosynthesis in the cytosol, the lipid-linked polysaccharide is transported to the periplasm by the WzmWzt ABC transporter. Often, O antigen secretion requires the chemical modification of its elongating terminus, which the transporter recognizes via a carbohydrate-binding domain (CBD). Here, using components from A. aeolicus, we identify the O antigen structure with methylated mannose or rhamnose as its cap. Crystal and cryo electron microscopy structures reveal how WzmWzt recognizes this cap between its carbohydrate and nucleotide-binding domains in a nucleotide-free state. ATP binding induces drastic conformational changes of its CBD, terminating interactions with the O antigen. ATPase assays and site directed mutagenesis reveal reduced hydrolytic activity upon O antigen binding, likely to facilitate polymer loading into the ABC transporter. Our results elucidate critical steps in the recognition and translocation of polysaccharides by ABC transporters. Nature Publishing Group UK 2022-09-05 /pmc/articles/PMC9445017/ /pubmed/36064941 http://dx.doi.org/10.1038/s41467-022-32597-2 Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Article Spellmon, Nicholas Muszyński, Artur Górniak, Ireneusz Vlach, Jiri Hahn, David Azadi, Parastoo Zimmer, Jochen Molecular basis for polysaccharide recognition and modulated ATP hydrolysis by the O antigen ABC transporter |
title | Molecular basis for polysaccharide recognition and modulated ATP hydrolysis by the O antigen ABC transporter |
title_full | Molecular basis for polysaccharide recognition and modulated ATP hydrolysis by the O antigen ABC transporter |
title_fullStr | Molecular basis for polysaccharide recognition and modulated ATP hydrolysis by the O antigen ABC transporter |
title_full_unstemmed | Molecular basis for polysaccharide recognition and modulated ATP hydrolysis by the O antigen ABC transporter |
title_short | Molecular basis for polysaccharide recognition and modulated ATP hydrolysis by the O antigen ABC transporter |
title_sort | molecular basis for polysaccharide recognition and modulated atp hydrolysis by the o antigen abc transporter |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9445017/ https://www.ncbi.nlm.nih.gov/pubmed/36064941 http://dx.doi.org/10.1038/s41467-022-32597-2 |
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