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Natural separation of the acyl-CoA ligase reaction results in a non-adenylating enzyme
Acyl-coenzyme A (CoA) ligases catalyze the activation of carboxylic acids via a two-step reaction of adenylation followed by thioesterification. Here, we report the discovery of a non-adenylating acyl-CoA ligase PtmA2 and the functional separation of an acyl-CoA ligase reaction. Both PtmA1 and PtmA2...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6008203/ https://www.ncbi.nlm.nih.gov/pubmed/29867143 http://dx.doi.org/10.1038/s41589-018-0061-0 |
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author | Wang, Nan Rudolf, Jeffrey D. Dong, Liao-Bin Osipiuk, Jerzy Hatzos-Skintges, Catherine Endres, Michael Chang, Chin-Yuan Babnigg, Gyorgy Joachimiak, Andrzej Phillips, George N. Shen, Ben |
author_facet | Wang, Nan Rudolf, Jeffrey D. Dong, Liao-Bin Osipiuk, Jerzy Hatzos-Skintges, Catherine Endres, Michael Chang, Chin-Yuan Babnigg, Gyorgy Joachimiak, Andrzej Phillips, George N. Shen, Ben |
author_sort | Wang, Nan |
collection | PubMed |
description | Acyl-coenzyme A (CoA) ligases catalyze the activation of carboxylic acids via a two-step reaction of adenylation followed by thioesterification. Here, we report the discovery of a non-adenylating acyl-CoA ligase PtmA2 and the functional separation of an acyl-CoA ligase reaction. Both PtmA1 and PtmA2, two acyl-CoA ligases from the biosynthetic pathway of platensimycin and platencin, are necessary for the two steps of CoA activation. Gene inactivation of ptmA1 and ptmA2 resulted in the accumulation of free acid and adenylate intermediates, respectively. Enzymatic and structural characterization of PtmA2 confirmed its ability to only catalyze thioesterification. Structural characterization of PtmA2 revealed it binds both free acid and adenylate substrates and undergoes the established mechanism of domain alternation. Finally, site-directed mutagenesis restored both the adenylation and complete CoA activation reactions. This study challenges the currently accepted paradigm of adenylating enzymes and inspires future investigations on functionally separated acyl-CoA ligases and their ramifications in biology. |
format | Online Article Text |
id | pubmed-6008203 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
record_format | MEDLINE/PubMed |
spelling | pubmed-60082032018-12-04 Natural separation of the acyl-CoA ligase reaction results in a non-adenylating enzyme Wang, Nan Rudolf, Jeffrey D. Dong, Liao-Bin Osipiuk, Jerzy Hatzos-Skintges, Catherine Endres, Michael Chang, Chin-Yuan Babnigg, Gyorgy Joachimiak, Andrzej Phillips, George N. Shen, Ben Nat Chem Biol Article Acyl-coenzyme A (CoA) ligases catalyze the activation of carboxylic acids via a two-step reaction of adenylation followed by thioesterification. Here, we report the discovery of a non-adenylating acyl-CoA ligase PtmA2 and the functional separation of an acyl-CoA ligase reaction. Both PtmA1 and PtmA2, two acyl-CoA ligases from the biosynthetic pathway of platensimycin and platencin, are necessary for the two steps of CoA activation. Gene inactivation of ptmA1 and ptmA2 resulted in the accumulation of free acid and adenylate intermediates, respectively. Enzymatic and structural characterization of PtmA2 confirmed its ability to only catalyze thioesterification. Structural characterization of PtmA2 revealed it binds both free acid and adenylate substrates and undergoes the established mechanism of domain alternation. Finally, site-directed mutagenesis restored both the adenylation and complete CoA activation reactions. This study challenges the currently accepted paradigm of adenylating enzymes and inspires future investigations on functionally separated acyl-CoA ligases and their ramifications in biology. 2018-06-04 2018-07 /pmc/articles/PMC6008203/ /pubmed/29867143 http://dx.doi.org/10.1038/s41589-018-0061-0 Text en Users may view, print, copy, and download text and data-mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use: http://www.nature.com/authors/editorial_policies/license.html#terms |
spellingShingle | Article Wang, Nan Rudolf, Jeffrey D. Dong, Liao-Bin Osipiuk, Jerzy Hatzos-Skintges, Catherine Endres, Michael Chang, Chin-Yuan Babnigg, Gyorgy Joachimiak, Andrzej Phillips, George N. Shen, Ben Natural separation of the acyl-CoA ligase reaction results in a non-adenylating enzyme |
title | Natural separation of the acyl-CoA ligase reaction results in a non-adenylating enzyme |
title_full | Natural separation of the acyl-CoA ligase reaction results in a non-adenylating enzyme |
title_fullStr | Natural separation of the acyl-CoA ligase reaction results in a non-adenylating enzyme |
title_full_unstemmed | Natural separation of the acyl-CoA ligase reaction results in a non-adenylating enzyme |
title_short | Natural separation of the acyl-CoA ligase reaction results in a non-adenylating enzyme |
title_sort | natural separation of the acyl-coa ligase reaction results in a non-adenylating enzyme |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6008203/ https://www.ncbi.nlm.nih.gov/pubmed/29867143 http://dx.doi.org/10.1038/s41589-018-0061-0 |
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