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The evolutionary advantage of an aromatic clamp in plant family 3 glycoside exo-hydrolases

In the barley β-d-glucan glucohydrolase, a glycoside hydrolase family 3 (GH3) enzyme, the Trp286/Trp434 clamp ensures β-d-glucosides binding, which is fundamental for substrate hydrolysis during plant growth and development. We employ mutagenesis, high-resolution X-ray crystallography, and multi-sca...

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Autores principales: Luang, Sukanya, Fernández-Luengo, Xavier, Nin-Hill, Alba, Streltsov, Victor A., Schwerdt, Julian G., Alonso-Gil, Santiago, Ketudat Cairns, James R., Pradeau, Stéphanie, Fort, Sébastien, Maréchal, Jean-Didier, Masgrau, Laura, Rovira, Carme, Hrmova, Maria
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9508125/
https://www.ncbi.nlm.nih.gov/pubmed/36151080
http://dx.doi.org/10.1038/s41467-022-33180-5
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author Luang, Sukanya
Fernández-Luengo, Xavier
Nin-Hill, Alba
Streltsov, Victor A.
Schwerdt, Julian G.
Alonso-Gil, Santiago
Ketudat Cairns, James R.
Pradeau, Stéphanie
Fort, Sébastien
Maréchal, Jean-Didier
Masgrau, Laura
Rovira, Carme
Hrmova, Maria
author_facet Luang, Sukanya
Fernández-Luengo, Xavier
Nin-Hill, Alba
Streltsov, Victor A.
Schwerdt, Julian G.
Alonso-Gil, Santiago
Ketudat Cairns, James R.
Pradeau, Stéphanie
Fort, Sébastien
Maréchal, Jean-Didier
Masgrau, Laura
Rovira, Carme
Hrmova, Maria
author_sort Luang, Sukanya
collection PubMed
description In the barley β-d-glucan glucohydrolase, a glycoside hydrolase family 3 (GH3) enzyme, the Trp286/Trp434 clamp ensures β-d-glucosides binding, which is fundamental for substrate hydrolysis during plant growth and development. We employ mutagenesis, high-resolution X-ray crystallography, and multi-scale molecular modelling methods to examine the binding and conformational behaviour of isomeric β-d-glucosides during substrate-product assisted processive catalysis that operates in GH3 hydrolases. Enzyme kinetics reveals that the W434H mutant retains broad specificity, while W434A behaves as a strict (1,3)-β-d-glucosidase. Investigations of reactant movements on the nanoscale reveal that processivity is sensitive to mutation-specific alterations of the tryptophan clamp. While wild-type and W434H utilise a lateral cavity for glucose displacement and sliding of (1,3)-linked hydrolytic products through the catalytic site without dissociation, consistent with their high hydrolytic rates, W434A does not adopt processive catalysis. Phylogenomic analyses of GH3 hydrolases disclose the evolutionary advantage of the tryptophan clamp that confers broad specificity, high catalytic efficiency, and processivity.
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spelling pubmed-95081252022-09-25 The evolutionary advantage of an aromatic clamp in plant family 3 glycoside exo-hydrolases Luang, Sukanya Fernández-Luengo, Xavier Nin-Hill, Alba Streltsov, Victor A. Schwerdt, Julian G. Alonso-Gil, Santiago Ketudat Cairns, James R. Pradeau, Stéphanie Fort, Sébastien Maréchal, Jean-Didier Masgrau, Laura Rovira, Carme Hrmova, Maria Nat Commun Article In the barley β-d-glucan glucohydrolase, a glycoside hydrolase family 3 (GH3) enzyme, the Trp286/Trp434 clamp ensures β-d-glucosides binding, which is fundamental for substrate hydrolysis during plant growth and development. We employ mutagenesis, high-resolution X-ray crystallography, and multi-scale molecular modelling methods to examine the binding and conformational behaviour of isomeric β-d-glucosides during substrate-product assisted processive catalysis that operates in GH3 hydrolases. Enzyme kinetics reveals that the W434H mutant retains broad specificity, while W434A behaves as a strict (1,3)-β-d-glucosidase. Investigations of reactant movements on the nanoscale reveal that processivity is sensitive to mutation-specific alterations of the tryptophan clamp. While wild-type and W434H utilise a lateral cavity for glucose displacement and sliding of (1,3)-linked hydrolytic products through the catalytic site without dissociation, consistent with their high hydrolytic rates, W434A does not adopt processive catalysis. Phylogenomic analyses of GH3 hydrolases disclose the evolutionary advantage of the tryptophan clamp that confers broad specificity, high catalytic efficiency, and processivity. Nature Publishing Group UK 2022-09-23 /pmc/articles/PMC9508125/ /pubmed/36151080 http://dx.doi.org/10.1038/s41467-022-33180-5 Text en © Crown 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
Luang, Sukanya
Fernández-Luengo, Xavier
Nin-Hill, Alba
Streltsov, Victor A.
Schwerdt, Julian G.
Alonso-Gil, Santiago
Ketudat Cairns, James R.
Pradeau, Stéphanie
Fort, Sébastien
Maréchal, Jean-Didier
Masgrau, Laura
Rovira, Carme
Hrmova, Maria
The evolutionary advantage of an aromatic clamp in plant family 3 glycoside exo-hydrolases
title The evolutionary advantage of an aromatic clamp in plant family 3 glycoside exo-hydrolases
title_full The evolutionary advantage of an aromatic clamp in plant family 3 glycoside exo-hydrolases
title_fullStr The evolutionary advantage of an aromatic clamp in plant family 3 glycoside exo-hydrolases
title_full_unstemmed The evolutionary advantage of an aromatic clamp in plant family 3 glycoside exo-hydrolases
title_short The evolutionary advantage of an aromatic clamp in plant family 3 glycoside exo-hydrolases
title_sort evolutionary advantage of an aromatic clamp in plant family 3 glycoside exo-hydrolases
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9508125/
https://www.ncbi.nlm.nih.gov/pubmed/36151080
http://dx.doi.org/10.1038/s41467-022-33180-5
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