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The crystal structure of CbpD clarifies substrate-specificity motifs in chitin-active lytic polysaccharide monooxygenases

Pseudomonas aeruginosa secretes diverse proteins via its type 2 secretion system, including a 39 kDa chitin-binding protein, CbpD. CbpD has recently been shown to be a lytic polysaccharide monooxygenase active on chitin and to contribute substantially to virulence. To date, no structure of this viru...

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Autores principales: Dade, Christopher M., Douzi, Badreddine, Cambillau, Christian, Ball, Genevieve, Voulhoux, Romé, Forest, Katrina T.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: International Union of Crystallography 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9344471/
https://www.ncbi.nlm.nih.gov/pubmed/35916229
http://dx.doi.org/10.1107/S2059798322007033
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author Dade, Christopher M.
Douzi, Badreddine
Cambillau, Christian
Ball, Genevieve
Voulhoux, Romé
Forest, Katrina T.
author_facet Dade, Christopher M.
Douzi, Badreddine
Cambillau, Christian
Ball, Genevieve
Voulhoux, Romé
Forest, Katrina T.
author_sort Dade, Christopher M.
collection PubMed
description Pseudomonas aeruginosa secretes diverse proteins via its type 2 secretion system, including a 39 kDa chitin-binding protein, CbpD. CbpD has recently been shown to be a lytic polysaccharide monooxygenase active on chitin and to contribute substantially to virulence. To date, no structure of this virulence factor has been reported. Its first two domains are homologous to those found in the crystal structure of Vibrio cholerae GbpA, while the third domain is homologous to the NMR structure of the CBM73 domain of Cellvibrio japonicus CjLPMO10A. Here, the 3.0 Å resolution crystal structure of CbpD solved by molecular replacement is reported, which required ab initio models of each CbpD domain generated by the artificial intelligence deep-learning structure-prediction algorithm RoseTTAFold. The structure of CbpD confirms some previously reported substrate-specificity motifs among LPMOAA10s, while challenging the predictive power of others. Additionally, the structure of CbpD shows that post-translational modifications occur on the chitin-binding surface. Moreover, the structure raises interesting possibilities about how type 2 secretion-system substrates may interact with the secretion machinery and demonstrates the utility of new artificial intelligence protein structure-prediction algorithms in making challenging structural targets tractable.
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spelling pubmed-93444712022-08-15 The crystal structure of CbpD clarifies substrate-specificity motifs in chitin-active lytic polysaccharide monooxygenases Dade, Christopher M. Douzi, Badreddine Cambillau, Christian Ball, Genevieve Voulhoux, Romé Forest, Katrina T. Acta Crystallogr D Struct Biol Research Papers Pseudomonas aeruginosa secretes diverse proteins via its type 2 secretion system, including a 39 kDa chitin-binding protein, CbpD. CbpD has recently been shown to be a lytic polysaccharide monooxygenase active on chitin and to contribute substantially to virulence. To date, no structure of this virulence factor has been reported. Its first two domains are homologous to those found in the crystal structure of Vibrio cholerae GbpA, while the third domain is homologous to the NMR structure of the CBM73 domain of Cellvibrio japonicus CjLPMO10A. Here, the 3.0 Å resolution crystal structure of CbpD solved by molecular replacement is reported, which required ab initio models of each CbpD domain generated by the artificial intelligence deep-learning structure-prediction algorithm RoseTTAFold. The structure of CbpD confirms some previously reported substrate-specificity motifs among LPMOAA10s, while challenging the predictive power of others. Additionally, the structure of CbpD shows that post-translational modifications occur on the chitin-binding surface. Moreover, the structure raises interesting possibilities about how type 2 secretion-system substrates may interact with the secretion machinery and demonstrates the utility of new artificial intelligence protein structure-prediction algorithms in making challenging structural targets tractable. International Union of Crystallography 2022-07-27 /pmc/articles/PMC9344471/ /pubmed/35916229 http://dx.doi.org/10.1107/S2059798322007033 Text en © Christopher M. Dade et al. 2022 https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution (CC-BY) Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original authors and source are cited.
spellingShingle Research Papers
Dade, Christopher M.
Douzi, Badreddine
Cambillau, Christian
Ball, Genevieve
Voulhoux, Romé
Forest, Katrina T.
The crystal structure of CbpD clarifies substrate-specificity motifs in chitin-active lytic polysaccharide monooxygenases
title The crystal structure of CbpD clarifies substrate-specificity motifs in chitin-active lytic polysaccharide monooxygenases
title_full The crystal structure of CbpD clarifies substrate-specificity motifs in chitin-active lytic polysaccharide monooxygenases
title_fullStr The crystal structure of CbpD clarifies substrate-specificity motifs in chitin-active lytic polysaccharide monooxygenases
title_full_unstemmed The crystal structure of CbpD clarifies substrate-specificity motifs in chitin-active lytic polysaccharide monooxygenases
title_short The crystal structure of CbpD clarifies substrate-specificity motifs in chitin-active lytic polysaccharide monooxygenases
title_sort crystal structure of cbpd clarifies substrate-specificity motifs in chitin-active lytic polysaccharide monooxygenases
topic Research Papers
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9344471/
https://www.ncbi.nlm.nih.gov/pubmed/35916229
http://dx.doi.org/10.1107/S2059798322007033
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