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Sets of Covariant Residues Modulate the Activity and Thermal Stability of GH1 β-Glucosidases
The statistical coupling analysis of 768 β-glucosidases from the GH1 family revealed 23 positions in which the amino acid frequencies are coupled. The roles of these covariant positions in terms of the properties of β-glucosidases were investigated by alanine-screening mutagenesis using the fall arm...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2014
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4013033/ https://www.ncbi.nlm.nih.gov/pubmed/24804841 http://dx.doi.org/10.1371/journal.pone.0096627 |
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author | Tamaki, Fábio K. Textor, Larissa C. Polikarpov, Igor Marana, Sandro R. |
author_facet | Tamaki, Fábio K. Textor, Larissa C. Polikarpov, Igor Marana, Sandro R. |
author_sort | Tamaki, Fábio K. |
collection | PubMed |
description | The statistical coupling analysis of 768 β-glucosidases from the GH1 family revealed 23 positions in which the amino acid frequencies are coupled. The roles of these covariant positions in terms of the properties of β-glucosidases were investigated by alanine-screening mutagenesis using the fall armyworm Spodoptera frugiperda β-glycosidase (Sfβgly) as a model. The effects of the mutations on the Sfβgly kinetic parameters (k (cat)/K (m)) for the hydrolysis of three different p-nitrophenyl β-glycosides and structural comparisons of several β-glucosidases showed that eleven covariant positions (54, 98, 143, 188, 195, 196, 203, 398, 451, 452 and 460 in Sfβgly numbering) form a layer surrounding the active site of the β-glucosidases, which modulates their catalytic activity and substrate specificity via direct contact with the active site residues. Moreover, the influence of the mutations on the transition temperature (T (m)) of Sfβgly indicated that nine of the coupled positions (49, 62, 143, 188, 223, 278, 309, 452 and 460 in Sfβgly numbering) are related to thermal stability. In addition to being preferentially occupied by prolines, structural comparisons indicated that these positions are concentrated at loop segments of the β-glucosidases. Therefore, due to these common biochemical and structural properties, these nine covariant positions, even without physical contacts among them, seem to jointly modulate the thermal stability of β-glucosidases. |
format | Online Article Text |
id | pubmed-4013033 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-40130332014-05-09 Sets of Covariant Residues Modulate the Activity and Thermal Stability of GH1 β-Glucosidases Tamaki, Fábio K. Textor, Larissa C. Polikarpov, Igor Marana, Sandro R. PLoS One Research Article The statistical coupling analysis of 768 β-glucosidases from the GH1 family revealed 23 positions in which the amino acid frequencies are coupled. The roles of these covariant positions in terms of the properties of β-glucosidases were investigated by alanine-screening mutagenesis using the fall armyworm Spodoptera frugiperda β-glycosidase (Sfβgly) as a model. The effects of the mutations on the Sfβgly kinetic parameters (k (cat)/K (m)) for the hydrolysis of three different p-nitrophenyl β-glycosides and structural comparisons of several β-glucosidases showed that eleven covariant positions (54, 98, 143, 188, 195, 196, 203, 398, 451, 452 and 460 in Sfβgly numbering) form a layer surrounding the active site of the β-glucosidases, which modulates their catalytic activity and substrate specificity via direct contact with the active site residues. Moreover, the influence of the mutations on the transition temperature (T (m)) of Sfβgly indicated that nine of the coupled positions (49, 62, 143, 188, 223, 278, 309, 452 and 460 in Sfβgly numbering) are related to thermal stability. In addition to being preferentially occupied by prolines, structural comparisons indicated that these positions are concentrated at loop segments of the β-glucosidases. Therefore, due to these common biochemical and structural properties, these nine covariant positions, even without physical contacts among them, seem to jointly modulate the thermal stability of β-glucosidases. Public Library of Science 2014-05-07 /pmc/articles/PMC4013033/ /pubmed/24804841 http://dx.doi.org/10.1371/journal.pone.0096627 Text en © 2014 Tamaki et al http://creativecommons.org/licenses/by/4.0/ 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 author and source are properly credited. |
spellingShingle | Research Article Tamaki, Fábio K. Textor, Larissa C. Polikarpov, Igor Marana, Sandro R. Sets of Covariant Residues Modulate the Activity and Thermal Stability of GH1 β-Glucosidases |
title | Sets of Covariant Residues Modulate the Activity and Thermal Stability of GH1 β-Glucosidases |
title_full | Sets of Covariant Residues Modulate the Activity and Thermal Stability of GH1 β-Glucosidases |
title_fullStr | Sets of Covariant Residues Modulate the Activity and Thermal Stability of GH1 β-Glucosidases |
title_full_unstemmed | Sets of Covariant Residues Modulate the Activity and Thermal Stability of GH1 β-Glucosidases |
title_short | Sets of Covariant Residues Modulate the Activity and Thermal Stability of GH1 β-Glucosidases |
title_sort | sets of covariant residues modulate the activity and thermal stability of gh1 β-glucosidases |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4013033/ https://www.ncbi.nlm.nih.gov/pubmed/24804841 http://dx.doi.org/10.1371/journal.pone.0096627 |
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