Morgan Charles W, Julien Olivier, Unger Elizabeth K, Shah Nirao M, Wells James A
Department of Pharmaceutical Chemistry, University of California, San Francisco, California, USA; Graduate Group in Chemistry and Chemical Biology, University of California, San Francisco, California, USA.
Department of Pharmaceutical Chemistry, University of California, San Francisco, California, USA.
Methods Enzymol. 2014;544:179-213. doi: 10.1016/B978-0-12-417158-9.00008-X.
Caspases, aspartate-specific cysteine proteases, have fate-determining roles in many cellular processes including apoptosis, differentiation, neuronal remodeling, and inflammation (for review, see Yuan & Kroemer, 2010). There are a dozen caspases in humans alone, yet their individual contributions toward these phenotypes are not well understood. Thus, there has been considerable interest in activating individual caspases or using their activity to drive these processes in cells and animals. We envision that such experimental control of caspase activity can not only afford novel insights into fundamental biological problems but may also enable new models for disease and suggest possible routes to therapeutic intervention. In particular, localized, genetic, and small-molecule-controlled caspase activation has the potential to target the desired cell type in a tissue. Suppression of caspase activation is one of the hallmarks of cancer and thus there has been significant enthusiasm for generating selective small-molecule activators that could bypass upstream mutational events that prevent apoptosis. Here, we provide a practical guide that investigators have devised, using genetics or small molecules, to activate specific caspases in cells or animals. Additionally, we show genetically controlled activation of an executioner caspase to target the function of a defined group of neurons in the adult mammalian brain.
半胱天冬酶，即天冬氨酸特异性半胱氨酸蛋白酶，在包括细胞凋亡、分化、神经元重塑和炎症在内的许多细胞过程中发挥着决定命运的作用（综述见Yuan & Kroemer，2010）。仅在人类中就有十几种半胱天冬酶，然而它们对这些表型的个体贡献尚未得到充分了解。因此，人们对激活单个半胱天冬酶或利用其活性在细胞和动物中驱动这些过程产生了浓厚兴趣。我们设想，对半胱天冬酶活性的这种实验性控制不仅可以为基本生物学问题提供新的见解，还可能为疾病建立新模型，并提出可能的治疗干预途径。特别是，局部、基因和小分子控制的半胱天冬酶激活有可能靶向组织中的所需细胞类型。半胱天冬酶激活的抑制是癌症的标志之一，因此人们对开发能够绕过阻止细胞凋亡的上游突变事件的选择性小分子激活剂有着极大的热情。在这里，我们提供了一份实用指南，介绍了研究人员利用遗传学或小分子在细胞或动物中激活特定半胱天冬酶的方法。此外，我们展示了通过基因控制激活一种执行性半胱天冬酶，以靶向成年哺乳动物大脑中一组特定神经元的功能。
