ABSTRACT
The AMP-forming acetyl-CoA synthetase is regulated by lysine acetylation both in bacteria and eukaryotes. However, the underlying mechanism is poorly understood. The Bacillus subtilis acetyltransferase AcuA and the AMP-forming acetyl-CoA synthetase AcsA form an AcuAâ¢AcsA complex, dissociating upon lysine acetylation of AcsA by AcuA. Crystal structures of AcsA from Chloroflexota bacterium in the apo form and in complex with acetyl-adenosine-5'-monophosphate (acetyl-AMP) support the flexible C-terminal domain adopting different conformations. AlphaFold2 predictions suggest binding of AcuA stabilizes AcsA in an undescribed conformation. We show the AcuAâ¢AcsA complex dissociates upon acetyl-coenzyme A (acetyl-CoA) dependent acetylation of AcsA by AcuA. We discover an intrinsic phosphotransacetylase activity enabling AcuAâ¢AcsA generating acetyl-CoA from acetyl-phosphate (AcP) and coenzyme A (CoA) used by AcuA to acetylate and inactivate AcsA. Here, we provide mechanistic insights into the regulation of AMP-forming acetyl-CoA synthetases by lysine acetylation and discover an intrinsic phosphotransacetylase allowing modulation of its activity based on AcP and CoA levels.
Subject(s)
Acetate-CoA Ligase , Acetyl Coenzyme A , Bacillus subtilis , Bacterial Proteins , Lysine , Acetylation , Lysine/metabolism , Acetyl Coenzyme A/metabolism , Acetate-CoA Ligase/metabolism , Acetate-CoA Ligase/genetics , Acetate-CoA Ligase/chemistry , Bacillus subtilis/metabolism , Bacillus subtilis/enzymology , Bacterial Proteins/metabolism , Bacterial Proteins/chemistry , Bacterial Proteins/genetics , Crystallography, X-Ray , Models, Molecular , Protein Binding , Adenosine Monophosphate/metabolism , OrganophosphatesABSTRACT
Biocatalysis has undergone revolutionary progress in the past century. Benefited by the integration of multidisciplinary technologies, natural enzymatic reactions are constantly being explored. Protein engineering gives birth to robust biocatalysts that are widely used in industrial production. These research achievements have gradually constructed a network containing natural enzymatic synthesis pathways and artificially designed enzymatic cascades. Nowadays, the development of artificial intelligence, automation, and ultra-high-throughput technology provides infinite possibilities for the discovery of novel enzymes, enzymatic mechanisms and enzymatic cascades, and gradually complements the lack of remaining key steps in the pathway design of enzymatic total synthesis. Therefore, the research of biocatalysis is gradually moving towards the era of novel technology integration, intelligent manufacturing and enzymatic total synthesis.
Subject(s)
Biocatalysis , Animals , Artificial Intelligence , Biosynthetic Pathways , Enzymes/metabolism , Humans , Protein EngineeringABSTRACT
This article presents a fair and balanced review of natural orifice translumenal endoscopic surgery. The article chronicles the history and technical aspects of natural orifice translumenal endoscopic surgery with particular emphasis on its application in urology. It is hoped that this article serves as a straightforward and pragmatic reference for practicing and academic urologists.
