Substituting the catalytic proline of 4-oxalocrotonate tautomerase with non-canonical analogues reveals a finely tuned catalytic system

The enzyme 4-oxalocrotonate tautomerase shows remarkable catalytic versatility due to the secondary amine of its N-terminal proline moiety. In this work, we incorporated a range of proline analogues into the enzyme and examined the effects on structure and activity. While the structure of the enzyme...

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Autores principales: Lukesch, Michael S., Pavkov-Keller, Tea, Gruber, Karl, Zangger, Klaus, Wiltschi, Birgit
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2019
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6389900/
https://www.ncbi.nlm.nih.gov/pubmed/30804446
http://dx.doi.org/10.1038/s41598-019-39484-9
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author Lukesch, Michael S.
Pavkov-Keller, Tea
Gruber, Karl
Zangger, Klaus
Wiltschi, Birgit
author_facet Lukesch, Michael S.
Pavkov-Keller, Tea
Gruber, Karl
Zangger, Klaus
Wiltschi, Birgit
author_sort Lukesch, Michael S.
collection PubMed
description The enzyme 4-oxalocrotonate tautomerase shows remarkable catalytic versatility due to the secondary amine of its N-terminal proline moiety. In this work, we incorporated a range of proline analogues into the enzyme and examined the effects on structure and activity. While the structure of the enzyme remained unperturbed, its promiscuous Michael-type activity was severely affected. This finding demonstrates how atomic changes in a biocatalytic system can abolish its activity. Our work provides a toolbox for successful generation of enzyme variants with non-canonical catalytic proline analogues.
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spelling pubmed-63899002019-02-28 Substituting the catalytic proline of 4-oxalocrotonate tautomerase with non-canonical analogues reveals a finely tuned catalytic system Lukesch, Michael S. Pavkov-Keller, Tea Gruber, Karl Zangger, Klaus Wiltschi, Birgit Sci Rep Article The enzyme 4-oxalocrotonate tautomerase shows remarkable catalytic versatility due to the secondary amine of its N-terminal proline moiety. In this work, we incorporated a range of proline analogues into the enzyme and examined the effects on structure and activity. While the structure of the enzyme remained unperturbed, its promiscuous Michael-type activity was severely affected. This finding demonstrates how atomic changes in a biocatalytic system can abolish its activity. Our work provides a toolbox for successful generation of enzyme variants with non-canonical catalytic proline analogues. Nature Publishing Group UK 2019-02-25 /pmc/articles/PMC6389900/ /pubmed/30804446 http://dx.doi.org/10.1038/s41598-019-39484-9 Text en © The Author(s) 2019 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/.
spellingShingle Article
Lukesch, Michael S.
Pavkov-Keller, Tea
Gruber, Karl
Zangger, Klaus
Wiltschi, Birgit
Substituting the catalytic proline of 4-oxalocrotonate tautomerase with non-canonical analogues reveals a finely tuned catalytic system
title Substituting the catalytic proline of 4-oxalocrotonate tautomerase with non-canonical analogues reveals a finely tuned catalytic system
title_full Substituting the catalytic proline of 4-oxalocrotonate tautomerase with non-canonical analogues reveals a finely tuned catalytic system
title_fullStr Substituting the catalytic proline of 4-oxalocrotonate tautomerase with non-canonical analogues reveals a finely tuned catalytic system
title_full_unstemmed Substituting the catalytic proline of 4-oxalocrotonate tautomerase with non-canonical analogues reveals a finely tuned catalytic system
title_short Substituting the catalytic proline of 4-oxalocrotonate tautomerase with non-canonical analogues reveals a finely tuned catalytic system
title_sort substituting the catalytic proline of 4-oxalocrotonate tautomerase with non-canonical analogues reveals a finely tuned catalytic system
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6389900/
https://www.ncbi.nlm.nih.gov/pubmed/30804446
http://dx.doi.org/10.1038/s41598-019-39484-9
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