Natural separation of the acyl-CoA ligase reaction results in a non-adenylating enzyme article

Nan Wang, Jeffrey D. Rudolf, Liao Bin Dong, Jerzy Osipiuk, Catherine Hatzos-Skintges, Michael Endres, Chin-Yuan Chang, Gyorgy Babnigg, Andrzej Joachimiak, George N. Phillips, Ben Shen

Research output: Contribution to journalArticlepeer-review

8 Scopus citations

Abstract

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.

Original languageEnglish
Pages (from-to)730-737
Number of pages8
JournalNature Chemical Biology
Volume14
Issue number7
DOIs
StatePublished - 1 Jul 2018

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