Chang David W, Yang Xiaolu
Abramson Family Cancer Research Institute and Department of Cancer Biology, University of Pennsylvania School of Medicine, Philadelphia, PA 19104, USA.
Sci STKE. 2003 Jan 28;2003(167):PL1. doi: 10.1126/stke.2003.167.pl1.
Oligomerization is an important biological mechanism for regulating signal transduction. Activation of caspases during apoptosis is triggered by adaptor protein-mediated oligomerization of initiator procaspases. To facilitate the study of initiator caspase activation, a system that allows inducible activation of various caspases both in vitro and in vivo is highly desired. Here we describe such a caspase activation system that is based on FK506 binding protein (FKBP)-mediated oligomerization. The NH(2)-terminal prodomains of initiator procaspases that facilitate the interaction between procaspases and their adaptor proteins are replaced by a derivative of FKBP called Fv. The Fv-caspase fusions can then be dimerized by a synthetic divalent Fv ligand, AP20187, which binds strongly to Fv but weakly to the endogenous FKBPs. This FKBP-based system may be widely applicable to the study of the regulation and functions of caspases.
寡聚化是调节信号转导的一种重要生物学机制。凋亡过程中半胱天冬酶的激活是由衔接蛋白介导的起始半胱天冬酶原寡聚化触发的。为便于研究起始半胱天冬酶的激活,非常需要一种能够在体外和体内诱导各种半胱天冬酶激活的系统。在此,我们描述了一种基于FK506结合蛋白(FKBP)介导的寡聚化的半胱天冬酶激活系统。起始半胱天冬酶原的NH(2)-末端前结构域促进半胱天冬酶原与其衔接蛋白之间的相互作用,被一种名为Fv的FKBP衍生物所取代。然后,Fv-半胱天冬酶融合蛋白可通过一种合成二价Fv配体AP20187二聚化,AP20187与Fv紧密结合,但与内源性FKBP结合较弱。这种基于FKBP的系统可能广泛应用于半胱天冬酶调节和功能的研究。