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A comparative structural analysis of the surface properties of asco-laccases
Laccases of different biological origins have been widely investigated and these studies have elucidated fundamentals of the generic catalytic mechanism. However, other features such as surface properties and residues located away from the catalytic centres may also have impact on enzyme function. H...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6218047/ https://www.ncbi.nlm.nih.gov/pubmed/30395580 http://dx.doi.org/10.1371/journal.pone.0206589 |
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author | Ernst, Heidi A. Jørgensen, Lise J. Bukh, Christian Piontek, Klaus Plattner, Dietmar A. Østergaard, Lars H. Larsen, Sine Bjerrum, Morten J. |
author_facet | Ernst, Heidi A. Jørgensen, Lise J. Bukh, Christian Piontek, Klaus Plattner, Dietmar A. Østergaard, Lars H. Larsen, Sine Bjerrum, Morten J. |
author_sort | Ernst, Heidi A. |
collection | PubMed |
description | Laccases of different biological origins have been widely investigated and these studies have elucidated fundamentals of the generic catalytic mechanism. However, other features such as surface properties and residues located away from the catalytic centres may also have impact on enzyme function. Here we present the crystal structure of laccase from Myceliophthora thermophila (MtL) to a resolution of 1.62 Å together with a thorough structural comparison with other members of the CAZy family AA1_3 that comprises fungal laccases from ascomycetes. The recombinant protein produced in A. oryzae has a molecular mass of 75 kDa, a pI of 4.2 and carries 13.5 kDa N-linked glycans. In the crystal, MtL forms a dimer with the phenolic substrate binding pocket blocked, suggesting that the active form of the enzyme is monomeric. Overall, the MtL structure conforms with the canonical fold of fungal laccases as well as the features specific for the asco-laccases. However, the structural comparisons also reveal significant variations within this taxonomic subgroup. Notable differences in the T1-Cu active site topology and polar motifs imply molecular evolution to serve different functional roles. Very few surface residues are conserved and it is noticeable that they encompass residues that interact with the N-glycans and/or are located at domain interfaces. The N-glycosylation sites are surprisingly conserved among asco-laccases and in most cases the glycan displays extensive interactions with the protein. In particular, the glycans at Asn88 and Asn210 appear to have evolved as an integral part of the asco-laccase structure. An uneven distribution of the carbohydrates around the enzyme give unique properties to a distinct part of the surface of the asco-laccases which may have implication for laccase function–in particular towards large substrates. |
format | Online Article Text |
id | pubmed-6218047 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-62180472018-11-19 A comparative structural analysis of the surface properties of asco-laccases Ernst, Heidi A. Jørgensen, Lise J. Bukh, Christian Piontek, Klaus Plattner, Dietmar A. Østergaard, Lars H. Larsen, Sine Bjerrum, Morten J. PLoS One Research Article Laccases of different biological origins have been widely investigated and these studies have elucidated fundamentals of the generic catalytic mechanism. However, other features such as surface properties and residues located away from the catalytic centres may also have impact on enzyme function. Here we present the crystal structure of laccase from Myceliophthora thermophila (MtL) to a resolution of 1.62 Å together with a thorough structural comparison with other members of the CAZy family AA1_3 that comprises fungal laccases from ascomycetes. The recombinant protein produced in A. oryzae has a molecular mass of 75 kDa, a pI of 4.2 and carries 13.5 kDa N-linked glycans. In the crystal, MtL forms a dimer with the phenolic substrate binding pocket blocked, suggesting that the active form of the enzyme is monomeric. Overall, the MtL structure conforms with the canonical fold of fungal laccases as well as the features specific for the asco-laccases. However, the structural comparisons also reveal significant variations within this taxonomic subgroup. Notable differences in the T1-Cu active site topology and polar motifs imply molecular evolution to serve different functional roles. Very few surface residues are conserved and it is noticeable that they encompass residues that interact with the N-glycans and/or are located at domain interfaces. The N-glycosylation sites are surprisingly conserved among asco-laccases and in most cases the glycan displays extensive interactions with the protein. In particular, the glycans at Asn88 and Asn210 appear to have evolved as an integral part of the asco-laccase structure. An uneven distribution of the carbohydrates around the enzyme give unique properties to a distinct part of the surface of the asco-laccases which may have implication for laccase function–in particular towards large substrates. Public Library of Science 2018-11-05 /pmc/articles/PMC6218047/ /pubmed/30395580 http://dx.doi.org/10.1371/journal.pone.0206589 Text en © 2018 Ernst 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 (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Article Ernst, Heidi A. Jørgensen, Lise J. Bukh, Christian Piontek, Klaus Plattner, Dietmar A. Østergaard, Lars H. Larsen, Sine Bjerrum, Morten J. A comparative structural analysis of the surface properties of asco-laccases |
title | A comparative structural analysis of the surface properties of asco-laccases |
title_full | A comparative structural analysis of the surface properties of asco-laccases |
title_fullStr | A comparative structural analysis of the surface properties of asco-laccases |
title_full_unstemmed | A comparative structural analysis of the surface properties of asco-laccases |
title_short | A comparative structural analysis of the surface properties of asco-laccases |
title_sort | comparative structural analysis of the surface properties of asco-laccases |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6218047/ https://www.ncbi.nlm.nih.gov/pubmed/30395580 http://dx.doi.org/10.1371/journal.pone.0206589 |
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