Dagbay Kevin, Eron Scott J, Serrano Banyuhay P, Velázquez-Delgado Elih M, Zhao Yunlong, Lin Di, Vaidya Sravanti, Hardy Jeanne A
Department of Chemistry, University of Massachusetts, Amherst, Massachusetts, USA.
Department of Chemistry, University of Massachusetts, Amherst, Massachusetts, USA.
Methods Enzymol. 2014;544:215-49. doi: 10.1016/B978-0-12-417158-9.00009-1.
One of the most promising and as yet underutilized means of regulating protein function is exploitation of allosteric sites. All caspases catalyze the same overall reaction, but they perform different biological roles and are differentially regulated. It is our hypothesis that many allosteric sites exist on various caspases and that understanding both the distinct and overlapping mechanisms by which each caspase can be allosterically controlled should ultimately enable caspase-specific inhibition. Here we describe the ongoing work and methods for compiling a comprehensive map of apoptotic caspase allostery. Central to this approach are the use of (i) the embedded record of naturally evolved allosteric sites that are sensitive to zinc-mediated inhibition, phosphorylation, and other posttranslational modifications, (ii) structural and mutagenic approaches, and (iii) novel binding sites identified by both rationally-designed and screening-derived small-molecule inhibitors.
调节蛋白质功能最有前景且尚未充分利用的方法之一是利用变构位点。所有半胱天冬酶催化相同的整体反应，但它们发挥不同的生物学作用且受到不同的调节。我们的假设是，各种半胱天冬酶上存在许多变构位点，并且了解每种半胱天冬酶可通过变构控制的独特和重叠机制最终应能实现半胱天冬酶特异性抑制。在此，我们描述了正在进行的工作以及编制凋亡半胱天冬酶变构综合图谱的方法。这种方法的核心是使用：（i）对锌介导的抑制、磷酸化和其他翻译后修饰敏感的自然进化变构位点的内在记录；（ii）结构和诱变方法；以及（iii）通过合理设计和筛选衍生的小分子抑制剂鉴定的新结合位点。
