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Crystal structure of the capsular polysaccharide synthesizing protein CapE of Staphylococcus aureus

Enzymes synthesizing the bacterial CP (capsular polysaccharide) are attractive antimicrobial targets. However, we lack critical information about the structure and mechanism of many of them. In an effort to reduce that gap, we have determined three different crystal structures of the enzyme CapE of...

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Autores principales: Miyafusa, Takamitsu, Caaveiro, Jose M. M., Tanaka, Yoshikazu, Tanner, Martin E., Tsumoto, Kouhei
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Portland Press Ltd. 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3699295/
https://www.ncbi.nlm.nih.gov/pubmed/23611437
http://dx.doi.org/10.1042/BSR20130017
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author Miyafusa, Takamitsu
Caaveiro, Jose M. M.
Tanaka, Yoshikazu
Tanner, Martin E.
Tsumoto, Kouhei
author_facet Miyafusa, Takamitsu
Caaveiro, Jose M. M.
Tanaka, Yoshikazu
Tanner, Martin E.
Tsumoto, Kouhei
author_sort Miyafusa, Takamitsu
collection PubMed
description Enzymes synthesizing the bacterial CP (capsular polysaccharide) are attractive antimicrobial targets. However, we lack critical information about the structure and mechanism of many of them. In an effort to reduce that gap, we have determined three different crystal structures of the enzyme CapE of the human pathogen Staphylococcus aureus. The structure reveals that CapE is a member of the SDR (short-chain dehydrogenase/reductase) super-family of proteins. CapE assembles in a hexameric complex stabilized by three major contact surfaces between protein subunits. Turnover of substrate and/or coenzyme induces major conformational changes at the contact interface between protein subunits, and a displacement of the substrate-binding domain with respect to the Rossmann domain. A novel dynamic element that we called the latch is essential for remodelling of the protein–protein interface. Structural and primary sequence alignment identifies a group of SDR proteins involved in polysaccharide synthesis that share the two salient features of CapE: the mobile loop (latch) and a distinctive catalytic site (MxxxK). The relevance of these structural elements was evaluated by site-directed mutagenesis.
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spelling pubmed-36992952013-07-08 Crystal structure of the capsular polysaccharide synthesizing protein CapE of Staphylococcus aureus Miyafusa, Takamitsu Caaveiro, Jose M. M. Tanaka, Yoshikazu Tanner, Martin E. Tsumoto, Kouhei Biosci Rep Original Paper Enzymes synthesizing the bacterial CP (capsular polysaccharide) are attractive antimicrobial targets. However, we lack critical information about the structure and mechanism of many of them. In an effort to reduce that gap, we have determined three different crystal structures of the enzyme CapE of the human pathogen Staphylococcus aureus. The structure reveals that CapE is a member of the SDR (short-chain dehydrogenase/reductase) super-family of proteins. CapE assembles in a hexameric complex stabilized by three major contact surfaces between protein subunits. Turnover of substrate and/or coenzyme induces major conformational changes at the contact interface between protein subunits, and a displacement of the substrate-binding domain with respect to the Rossmann domain. A novel dynamic element that we called the latch is essential for remodelling of the protein–protein interface. Structural and primary sequence alignment identifies a group of SDR proteins involved in polysaccharide synthesis that share the two salient features of CapE: the mobile loop (latch) and a distinctive catalytic site (MxxxK). The relevance of these structural elements was evaluated by site-directed mutagenesis. Portland Press Ltd. 2013-06-11 /pmc/articles/PMC3699295/ /pubmed/23611437 http://dx.doi.org/10.1042/BSR20130017 Text en © 2013 The author(s) has paid for this article to be freely available under the terms of the Creative Commons Attribution Licence (CC-BY)(http://creativecommons.org/licenses/by/3.0/) which permits unrestricted use, distribution and reproduction in any medium, provided the original work is properly cited. http://creativecommons.org/licenses/by/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Original Paper
Miyafusa, Takamitsu
Caaveiro, Jose M. M.
Tanaka, Yoshikazu
Tanner, Martin E.
Tsumoto, Kouhei
Crystal structure of the capsular polysaccharide synthesizing protein CapE of Staphylococcus aureus
title Crystal structure of the capsular polysaccharide synthesizing protein CapE of Staphylococcus aureus
title_full Crystal structure of the capsular polysaccharide synthesizing protein CapE of Staphylococcus aureus
title_fullStr Crystal structure of the capsular polysaccharide synthesizing protein CapE of Staphylococcus aureus
title_full_unstemmed Crystal structure of the capsular polysaccharide synthesizing protein CapE of Staphylococcus aureus
title_short Crystal structure of the capsular polysaccharide synthesizing protein CapE of Staphylococcus aureus
title_sort crystal structure of the capsular polysaccharide synthesizing protein cape of staphylococcus aureus
topic Original Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3699295/
https://www.ncbi.nlm.nih.gov/pubmed/23611437
http://dx.doi.org/10.1042/BSR20130017
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