Eukaryotic arylamine N-acetyltransferase investigation of substrate specificity by high-throughput screening

被引：73

作者：

Kawamura, A

Graham, J

Mushtaq, A

Tsiftsoglou, SA

Vath, GM

Hanna, PE

Wagner, CR

Sim, E

机构：

[1] Univ Oxford, Dept Pharmacol, Oxford OX1 3QT, England

[2] Univ Oxford, Dept Biochem, Oxford OX1 3QU, England

[3] Univ Minnesota, Dept Med Chem, Minneapolis, MN 55455 USA

[4] Univ Minnesota, Dept Pharmacol, Minneapolis, MN 55455 USA

来源：

BIOCHEMICAL PHARMACOLOGY | 2005年 / 69卷 / 02期

基金：

英国惠康基金;

关键词：

arylamine N-acetyltransferase; xenobiotic metabolism; recombinant protein; high-throughput screening; arylamines; acetylCoA/CoA derivatives;

D O I：

10.1016/j.bcp.2004.09.014

中图分类号：

R9 [药学];

学科分类号：

1007 ;

摘要：

Arylamine N-acetyltransferases (NAT; EC 2.3.1.5) catalyse the transfer of acetyl groups from acetylCoA to xenobiotics, including drugs and carcinogens. The enzyme is found extensively in both eukaryotes and prokaryotes, yet the endogenous roles of NATs are still unclear. In order to study the properties of eukaryotic NATs, high-throughput substrate and inhibitor screens have been developed using pure soluble recombinant Syrian hamster NAT2 (shNAT2) protein. The assay can be used with a wide range of compounds and was used to determine substrate specificity of shNAT2. We describe the expression and characterisation of shNAT2 and also purified recombinant human NAT1 and NAT2, including the use of the assay to explore the substrate specificities of each of the enzymes. Hamster NAT2 has similar substrate specificity to human NAT1, acetylating para-aminobenzoate but not arylhydrazine and hydralazine compounds. The overlapping but distinct substrate-specific activity profiles of human NAT1 and NAT2 were clearly observed from the screen. Naturally occurring compounds were tested as substrates or inhibitors of shNAT2 and succinylCoA was found to be a potent inhibitor of shNAT2. (C) 2004 Elsevier Inc. All rights reserved.

引用

页码：347 / 359

页数：13

共 57 条

[11] DRUG-METABOLIZING N-ACETYLTRANSFERASE ACTIVITY IN HUMAN CELL-LINES [J].