氯霉素乙酰转移酶III和大肠杆菌β-酮酰合酶III与部分水解的乙酰氧(dethia)辅酶a共结晶的结构

IF 1.1 4区 生物学 Q4 BIOCHEMICAL RESEARCH METHODS Acta crystallographica. Section F, Structural biology communications Pub Date : 2023-02-23 DOI:10.1107/S2053230X23001206
Aaron B. Benjamin, Lee M. Stunkard, Jianheng Ling, Jaelen N. Nice, Jeremy R. Lohman
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引用次数: 0

摘要

乙酰辅酶A(Acetyl-CoA)是一种反应性代谢产物,在结晶时间内在许多酶活性位点进行非生产性水解。为了阐明导致催化的酶-乙酰辅酶A相互作用,需要乙酰辅酶A底物类似物。在结构研究中使用的一种可能的类似物是乙酰氧杂(dethia)CoA(AcOCoA),其中CoA的硫酯S原子被O原子取代。本文介绍了在部分水解的AcOCoA和各自亲核试剂存在下生长的晶体中的氯霉素乙酰转移酶III(CATIII)和大肠杆菌酮酰基合酶III(FabH)的结构。根据结构,AcOCoA在不同酶之间的行为不同,FabH与AcOCoA反应,CATIII不起作用。CATIII的结构揭示了对催化机制的深入了解,三聚物的一个活性位点对AcOCoA和氯霉素具有相对清晰的电子密度,而其他活性位点对AcOCoA具有较弱的密度。一个FabH结构包含水解的AcOCoA产物oxa(dethia)CoA(OCoA),而另一个FabH结构包含具有OCoA的酰基酶中间体。总之,这些结构为AcOCoA用于不同亲核试剂的酶结构-功能研究提供了初步见解。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

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Structures of chloramphenicol acetyltransferase III and Escherichia coli β-ketoacylsynthase III co-crystallized with partially hydrolysed acetyl-oxa(dethia)CoA

Acetyl coenzyme A (acetyl-CoA) is a reactive metabolite that nonproductively hydrolyzes in a number of enzyme active sites in the crystallization time frame. In order to elucidate the enzyme–acetyl-CoA interactions leading to catalysis, acetyl-CoA substrate analogs are needed. One possible analog for use in structural studies is acetyl-oxa(dethia)CoA (AcOCoA), in which the thioester S atom of CoA is replaced by an O atom. Here, structures of chloramphenicol acetyltransferase III (CATIII) and Escherichia coli ketoacylsynthase III (FabH) from crystals grown in the presence of partially hydrolyzed AcOCoA and the respective nucleophile are presented. Based on the structures, the behavior of AcOCoA differs between the enzymes, with FabH reacting with AcOCoA and CATIII being unreactive. The structure of CATIII reveals insight into the catalytic mechanism, with one active site of the trimer having relatively clear electron density for AcOCoA and chloramphenicol and the other active sites having weaker density for AcOCoA. One FabH structure contains a hydrolyzed AcOCoA product oxa(dethia)CoA (OCoA), while the other FabH structure contains an acyl-enzyme intermediate with OCoA. Together, these structures provide preliminary insight into the use of AcOCoA for enzyme structure–function studies with different nucleophiles.

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来源期刊
Acta crystallographica. Section F, Structural biology communications
Acta crystallographica. Section F, Structural biology communications BIOCHEMICAL RESEARCH METHODSBIOCHEMISTRY &-BIOCHEMISTRY & MOLECULAR BIOLOGY
CiteScore
1.90
自引率
0.00%
发文量
95
期刊介绍: Acta Crystallographica Section F is a rapid structural biology communications journal. Articles on any aspect of structural biology, including structures determined using high-throughput methods or from iterative studies such as those used in the pharmaceutical industry, are welcomed by the journal. The journal offers the option of open access, and all communications benefit from unlimited free use of colour illustrations and no page charges. Authors are encouraged to submit multimedia content for publication with their articles. Acta Cryst. F has a dedicated online tool called publBio that is designed to make the preparation and submission of articles easier for authors.
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