Bm-iAANAT3: Expression and characterization of a novel arylalkylamine N-acyltransferase from Bombyx mori
| Autor: | Brian G. O'Flynn, Lamar Galloway, Christopher Shoji, David J. Merkler, Matthew R. Battistini, Gabriela Suarez |
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| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
Stereochemistry Biophysics Gene Expression Arylalkylamine N-Acetyltransferase Biochemistry Article Substrate Specificity 03 medical and health sciences Bombyx mori Animals Molecular Biology chemistry.chemical_classification 030102 biochemistry & molecular biology biology Mutagenesis Acetylation Bombyx biology.organism_classification Recombinant Proteins Amino acid Kinetics 030104 developmental biology Enzyme chemistry Acyltransferase Arylalkylamine Insect Proteins lipids (amino acids peptides and proteins) Amine gas treating |
| Zdroj: | Archives of Biochemistry and Biophysics. 661:107-116 |
| ISSN: | 0003-9861 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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