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Site-specific bioalkylation of rapamycin by the RapM 16-O-methyltransferase

The methylation of natural products by S-adenosyl methionine (AdoMet, also known as SAM)-dependent methyltransferase enzymes is a common tailoring step in many biosynthetic pathways. The introduction of methyl substituents can affect the biological and physicochemical properties of the secondary met...

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Autores principales: Law, Brian J. C., Struck, Anna-Winona, Bennett, Matthew R., Wilkinson, Barrie, Micklefield, Jason
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Royal Society of Chemistry 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5729408/
https://www.ncbi.nlm.nih.gov/pubmed/29403635
http://dx.doi.org/10.1039/c5sc00164a
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author Law, Brian J. C.
Struck, Anna-Winona
Bennett, Matthew R.
Wilkinson, Barrie
Micklefield, Jason
author_facet Law, Brian J. C.
Struck, Anna-Winona
Bennett, Matthew R.
Wilkinson, Barrie
Micklefield, Jason
author_sort Law, Brian J. C.
collection PubMed
description The methylation of natural products by S-adenosyl methionine (AdoMet, also known as SAM)-dependent methyltransferase enzymes is a common tailoring step in many biosynthetic pathways. The introduction of methyl substituents can affect the biological and physicochemical properties of the secondary metabolites produced. Recently it has become apparent that some AdoMet-dependent methyltransferases exhibit promiscuity and will accept AdoMet analogues enabling the transfer of alternative alkyl groups. In this study we have characterised a methyltransferase, RapM, which is involved in the biosynthesis of the potent immunosuppressive agent rapamycin. We have shown that recombinant RapM regioselectively methylates the C16 hydroxyl group of desmethyl rapamycin precursors in vitro and is promiscuous in accepting alternative co-factors in addition to AdoMet. A coupled enzyme system was developed, including a mutant human enzyme methionine adenosyl transferase (MAT), along with RapM, which was used to prepare alkylated rapamycin derivatives (rapalogs) with alternative ethyl and allyl ether groups, derived from simple S-ethyl or S-allyl methionine analogues. There are two other methyltransferases RapI and RapQ which provide methyl substituents of rapamycin. Consequently, using the enzymatic approach described here, it should be possible to generate a diverse array of alkylated rapalogs, with altered properties, that would be difficult to obtain by traditional synthetic approaches.
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spelling pubmed-57294082018-02-05 Site-specific bioalkylation of rapamycin by the RapM 16-O-methyltransferase Law, Brian J. C. Struck, Anna-Winona Bennett, Matthew R. Wilkinson, Barrie Micklefield, Jason Chem Sci Chemistry The methylation of natural products by S-adenosyl methionine (AdoMet, also known as SAM)-dependent methyltransferase enzymes is a common tailoring step in many biosynthetic pathways. The introduction of methyl substituents can affect the biological and physicochemical properties of the secondary metabolites produced. Recently it has become apparent that some AdoMet-dependent methyltransferases exhibit promiscuity and will accept AdoMet analogues enabling the transfer of alternative alkyl groups. In this study we have characterised a methyltransferase, RapM, which is involved in the biosynthesis of the potent immunosuppressive agent rapamycin. We have shown that recombinant RapM regioselectively methylates the C16 hydroxyl group of desmethyl rapamycin precursors in vitro and is promiscuous in accepting alternative co-factors in addition to AdoMet. A coupled enzyme system was developed, including a mutant human enzyme methionine adenosyl transferase (MAT), along with RapM, which was used to prepare alkylated rapamycin derivatives (rapalogs) with alternative ethyl and allyl ether groups, derived from simple S-ethyl or S-allyl methionine analogues. There are two other methyltransferases RapI and RapQ which provide methyl substituents of rapamycin. Consequently, using the enzymatic approach described here, it should be possible to generate a diverse array of alkylated rapalogs, with altered properties, that would be difficult to obtain by traditional synthetic approaches. Royal Society of Chemistry 2015-05-01 2015-03-02 /pmc/articles/PMC5729408/ /pubmed/29403635 http://dx.doi.org/10.1039/c5sc00164a Text en This journal is © The Royal Society of Chemistry 2015 http://creativecommons.org/licenses/by/3.0/ This article is freely available. This article is licensed under a Creative Commons Attribution 3.0 Unported Licence (CC BY 3.0)
spellingShingle Chemistry
Law, Brian J. C.
Struck, Anna-Winona
Bennett, Matthew R.
Wilkinson, Barrie
Micklefield, Jason
Site-specific bioalkylation of rapamycin by the RapM 16-O-methyltransferase
title Site-specific bioalkylation of rapamycin by the RapM 16-O-methyltransferase
title_full Site-specific bioalkylation of rapamycin by the RapM 16-O-methyltransferase
title_fullStr Site-specific bioalkylation of rapamycin by the RapM 16-O-methyltransferase
title_full_unstemmed Site-specific bioalkylation of rapamycin by the RapM 16-O-methyltransferase
title_short Site-specific bioalkylation of rapamycin by the RapM 16-O-methyltransferase
title_sort site-specific bioalkylation of rapamycin by the rapm 16-o-methyltransferase
topic Chemistry
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5729408/
https://www.ncbi.nlm.nih.gov/pubmed/29403635
http://dx.doi.org/10.1039/c5sc00164a
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