Democritus University of Thrace, Department of Molecular Biology and Genetics, Alexandroupolis, Greece.
IRD UMR216, Mère et enfant face aux infections tropicales, Paris, France.
Sci Rep. 2018 Jun 27;8(1):9759. doi: 10.1038/s41598-018-28094-6.
Xenobiotic metabolising N-acetyltransferases (NATs) perform biotransformation of drugs and carcinogens. Human NAT1 is associated with endogenous metabolic pathways of cells and is a candidate drug target for cancer. Human NAT2 is a well-characterised polymorphic xenobiotic metabolising enzyme, modulating susceptibility to drug-induced toxicity. Human NATs are difficult to express to high purification yields, complicating large-scale production for high-throughput screens or use in sophisticated enzymology assays and crystallography. We undertake comparative functional investigation of the NAT homologues of ten non-human primates, to characterise their properties and evaluate their suitability as models of human NATs. Considering the amount of generated recombinant protein, the enzymatic activity and thermal stability, the NAT homologues of non-human primates are demonstrated to be a much more effective resource for in vitro studies compared with human NATs. Certain NAT homologues are proposed as better models, such as the NAT1 of macaques Macaca mulatta and M. sylvanus, the NAT2 of Erythrocebus patas, and both NAT proteins of the gibbon Nomascus gabriellae which show highest homology to human NATs. This comparative investigation will facilitate in vitro screens towards discovery and optimisation of candidate pharmaceutical compounds for human NAT isoenzymes, while enabling better understanding of NAT function and evolution in primates.
异源生物代谢 N-乙酰基转移酶（NATs）对药物和致癌物质进行生物转化。人类 NAT1 与细胞内源性代谢途径有关，是癌症药物靶点的候选物。人类 NAT2 是一种特征明确的多态性异源生物代谢酶，调节对药物诱导毒性的易感性。人类 NATs 难以高纯度产量表达，这使得大规模生产用于高通量筛选或在复杂的酶学测定和晶体学中使用变得复杂。我们对十种非人类灵长类动物的 NAT 同源物进行了比较功能研究，以表征它们的特性，并评估它们作为人类 NAT 模型的适用性。考虑到产生的重组蛋白的量、酶活性和热稳定性，与人类 NATs 相比，非人类灵长类动物的 NAT 同源物被证明是更有效的体外研究资源。某些 NAT 同源物被提议作为更好的模型，例如猕猴 Macaca mulatta 和 M. sylvanus 的 NAT1、Erythrocebus patas 的 NAT2 以及 Nomascus gabriellae 的两种 gibbon NAT 蛋白，它们与人 NATs 的同源性最高。这种比较研究将有助于对人类 NAT 同工酶的候选药物化合物进行体外筛选，以发现和优化，同时使人们更好地了解灵长类动物中 NAT 的功能和进化。
