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Evolution of the key alkaloid enzyme putrescine N-methyltransferase from spermidine synthase
Putrescine N-methyltransferases (PMTs) are the first specific enzymes of the biosynthesis of nicotine and tropane alkaloids. PMTs transfer a methyl group onto the diamine putrescine from S-adenosyl-l-methionine (SAM) as coenzyme. PMT proteins have presumably evolved from spermidine synthases (SPDSs)...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Frontiers Media S.A.
2013
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3725402/ https://www.ncbi.nlm.nih.gov/pubmed/23908659 http://dx.doi.org/10.3389/fpls.2013.00260 |
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author | Junker, Anne Fischer, Juliane Sichhart, Yvonne Brandt, Wolfgang Dräger, Birgit |
author_facet | Junker, Anne Fischer, Juliane Sichhart, Yvonne Brandt, Wolfgang Dräger, Birgit |
author_sort | Junker, Anne |
collection | PubMed |
description | Putrescine N-methyltransferases (PMTs) are the first specific enzymes of the biosynthesis of nicotine and tropane alkaloids. PMTs transfer a methyl group onto the diamine putrescine from S-adenosyl-l-methionine (SAM) as coenzyme. PMT proteins have presumably evolved from spermidine synthases (SPDSs), which are ubiquitous enzymes of polyamine metabolism. SPDSs use decarboxylated SAM as coenzyme to transfer an aminopropyl group onto putrescine. In an attempt to identify possible and necessary steps in the evolution of PMT from SPDS, homology based modeling of Datura stramonium SPDS1 and PMT was employed to gain deeper insight in the preferred binding positions and conformations of the substrate and the alternative coenzymes. Based on predictions of amino acids responsible for the change of enzyme specificities, sites of mutagenesis were derived. PMT activity was generated in D. stramonium SPDS1 after few amino acid exchanges. Concordantly, Arabidopsis thaliana SPDS1 was mutated and yielded enzymes with both, PMT and SPDS activities. Kinetic parameters were measured for enzymatic characterization. The switch from aminopropyl to methyl transfer depends on conformational changes of the methionine part of the coenzyme in the binding cavity of the enzyme. The rapid generation of PMT activity in SPDS proteins and the wide-spread occurrence of putative products of N-methylputrescine suggest that PMT activity is present frequently in the plant kingdom. |
format | Online Article Text |
id | pubmed-3725402 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-37254022013-08-01 Evolution of the key alkaloid enzyme putrescine N-methyltransferase from spermidine synthase Junker, Anne Fischer, Juliane Sichhart, Yvonne Brandt, Wolfgang Dräger, Birgit Front Plant Sci Plant Science Putrescine N-methyltransferases (PMTs) are the first specific enzymes of the biosynthesis of nicotine and tropane alkaloids. PMTs transfer a methyl group onto the diamine putrescine from S-adenosyl-l-methionine (SAM) as coenzyme. PMT proteins have presumably evolved from spermidine synthases (SPDSs), which are ubiquitous enzymes of polyamine metabolism. SPDSs use decarboxylated SAM as coenzyme to transfer an aminopropyl group onto putrescine. In an attempt to identify possible and necessary steps in the evolution of PMT from SPDS, homology based modeling of Datura stramonium SPDS1 and PMT was employed to gain deeper insight in the preferred binding positions and conformations of the substrate and the alternative coenzymes. Based on predictions of amino acids responsible for the change of enzyme specificities, sites of mutagenesis were derived. PMT activity was generated in D. stramonium SPDS1 after few amino acid exchanges. Concordantly, Arabidopsis thaliana SPDS1 was mutated and yielded enzymes with both, PMT and SPDS activities. Kinetic parameters were measured for enzymatic characterization. The switch from aminopropyl to methyl transfer depends on conformational changes of the methionine part of the coenzyme in the binding cavity of the enzyme. The rapid generation of PMT activity in SPDS proteins and the wide-spread occurrence of putative products of N-methylputrescine suggest that PMT activity is present frequently in the plant kingdom. Frontiers Media S.A. 2013-07-29 /pmc/articles/PMC3725402/ /pubmed/23908659 http://dx.doi.org/10.3389/fpls.2013.00260 Text en Copyright © 2013 Junker, Fischer, Sichhart, Brandt and Dräger. http://creativecommons.org/licenses/by/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in other forums, provided the original authors and source are credited and subject to any copyright notices concerning any third-party graphics etc. |
spellingShingle | Plant Science Junker, Anne Fischer, Juliane Sichhart, Yvonne Brandt, Wolfgang Dräger, Birgit Evolution of the key alkaloid enzyme putrescine N-methyltransferase from spermidine synthase |
title | Evolution of the key alkaloid enzyme putrescine N-methyltransferase from spermidine synthase |
title_full | Evolution of the key alkaloid enzyme putrescine N-methyltransferase from spermidine synthase |
title_fullStr | Evolution of the key alkaloid enzyme putrescine N-methyltransferase from spermidine synthase |
title_full_unstemmed | Evolution of the key alkaloid enzyme putrescine N-methyltransferase from spermidine synthase |
title_short | Evolution of the key alkaloid enzyme putrescine N-methyltransferase from spermidine synthase |
title_sort | evolution of the key alkaloid enzyme putrescine n-methyltransferase from spermidine synthase |
topic | Plant Science |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3725402/ https://www.ncbi.nlm.nih.gov/pubmed/23908659 http://dx.doi.org/10.3389/fpls.2013.00260 |
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