Structural Study of Sulfotransferases

工作总结：磺基转移酶代谢是一种生物信号如类固醇激素的失活，也是一种防御系统，对环境毒物和致癌物。为了遇到几乎无限数量的结构多样的化学物质，这些酶表现出广泛的底物特异性。因此，重要的是要研究决定底物特异性的结构原理，以预测代谢和由此产生的后果，人类易感性的化学品暴露。对于调查，我们使用了X射线晶体学和定点诱变。我们解决了雌激素磺基转移酶（EST）与PAP和雌二醇以及与PAP和钒酸盐复合的晶体结构。这些结构为深入理解磺基转移酶催化的反应机理提供了依据。基于EST底物口袋中的狭窄样门结构的定点突变赋予酶雌激素特异性。- 磺基转移酶，x射线晶体学，酶作用，定点突变，蛋白质表达和纯化

Summary of Work: Metabolism by sulfotransferases is an inactivation of biological signals such as steroid hormones and also a defense system against environmental toxicants and carcinogens. To encounter virtually unlimited numbers of structurally diverse chemiclas, these enzymes exhibit broad substrate specificity. Thus, it is important to investigate the structural principle determining the substrate specificity in order to predict the metabolisms and the resulted consequences to human susceptibililty to chemical exposure. For the investigation, we have used x-ray crystallography and site-directed mutagenesis. We have solved the crystal structure of estrogen sulfotransferase (EST) complexed with PAP and estradiol and that with PAP and vanadate. These structures provided the insight into understanding the reaction mechanism catalyzed by sulfotransferase. Site-directed mutagenesis based on the structres defined the stricture- like gate in the substrate pocket of EST that confers estrogen specificity to the enzyme. - Sulfotranserase, x-ray crystallography, enzyme action, site-directed mutagenesis, protein expression and purification