家蚕新型芳基烷基胺 N-酰基转移酶的表达与特性研究。
Bm-iAANAT3: Expression and characterization of a novel arylalkylamine N-acyltransferase from Bombyx mori.
机构信息
Department of Chemistry, University of South Florida, Tampa, FL, 33620, USA.
Department of Chemistry, University of South Florida, Tampa, FL, 33620, USA.
出版信息
Arch Biochem Biophys. 2019 Jan;661:107-116. doi: 10.1016/j.abb.2018.11.015. Epub 2018 Nov 16.
The arylalkylamine N-acyltransferases (AANATs) are enzymes that catalyze the acyl-CoA-dependent formation of N-acylarylalkylamides: acyl-CoA + arylalkylamine → N-acylarylalkylamides + CoA-SH. Herein, we describe our study of a previously uncharacterized AANAT from Bombyx mori: Bm-iAANAT3. Bm-iAANAT3 catalyzes the direct formation of N-acylarylalkylamides and accepts a broad range of short-chain acyl-CoA thioesters and amines as substrates. Acyl-CoA thioesters possessing an acyl chain length >10 carbon atoms are not substrates for Bm-iAANAT3. We report that Bm-iAANAT3 is a "versatile generalist", most likely, functioning in amine acetylation - a reaction in amine inactivation/excretion, cuticle sclerotization, and melanism. We propose a kinetic and chemical mechanism for Bm-iAANAT3 that is consistent with our steady-state kinetic analysis, dead-end inhibition studies, determination of the pH-rate profiles, and site-directed mutagenesis of a catalytically important amino acid in Bm-iAANAT3. These mechanistic studies of Bm-iAANAT3 will foster the development of novel compounds targeted against this enzyme and other insect AANATs for the control of insect pests.
芳基烷基胺 N-酰基转移酶（AANATs）是一种酶，可催化酰基辅酶 A 依赖性形成 N-酰基芳基烷基酰胺：酰基辅酶 A + 芳基烷基胺 → N-酰基芳基烷基酰胺 + 辅酶 A-SH。在此，我们描述了对来自家蚕的先前未表征的 AANAT 的研究：Bm-iAANAT3。Bm-iAANAT3 催化 N-酰基芳基烷基酰胺的直接形成，并接受广泛的短链酰基辅酶 A 硫酯和胺作为底物。酰基辅酶 A 硫酯的酰链长度> 10 个碳原子不是 Bm-iAANAT3 的底物。我们报告说，Bm-iAANAT3 是一种“通用多面手”，很可能在胺乙酰化 - 一种胺失活/排泄、角质层硬化和黑化的反应中发挥作用。我们提出了一个符合我们稳态动力学分析、末端抑制研究、pH-速率曲线测定和 Bm-iAANAT3 中催化重要氨基酸的定点突变的 Bm-iAANAT3 的动力学和化学机制。对 Bm-iAANAT3 的这些机制研究将促进针对该酶和其他昆虫 AANAT 的新型化合物的开发，以控制昆虫害虫。
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