Park Sun Ah, Shaked Gideon M, Bredesen Dale E, Koo Edward H
Department of Neurosciences, University of California, San Diego, La Jolla, CA 92093, USA.
Biochem Biophys Res Commun. 2009 Oct 16;388(2):450-5. doi: 10.1016/j.bbrc.2009.08.042. Epub 2009 Aug 11.
The cytoplasmic tail of the amyloid precursor protein (APP) contains two putatively cytotoxic peptides, Jcasp and C31, derived by caspase cleavage of APP. Jcasp is a fragment starting from the epsilon-secretase site to position 664, while C31 is a fragment from position 665 to the C-terminus. Our studies now showed that compared to C31, Jcasp appeared to play a minor role in cytotoxicity. In particular, inhibition of Jcasp generation by treatment of gamma-secretase inhibitor did not lead to any attenuation of C31-induced toxicity. Secondly, because C31 toxicity is largely absent in cells lacking endogenous APP, we determined, using a split beta-galactosidase complementary assay to monitor protein-protein interactions, the presence of APP associated complexes. Our results demonstrated that both APP homomeric and C31/APP heteromeric complexes were correlated with cell death, indicating that C31 complexes with APP to recruit the interacting partners that initiate the signals related to cellular toxicity.
淀粉样前体蛋白(APP)的胞质尾部包含两种可能具有细胞毒性的肽段,即Jcasp和C31,它们是由APP经半胱天冬酶切割产生的。Jcasp是一个从ε-分泌酶切割位点开始至第664位的片段,而C31是一个从第665位至C末端的片段。我们目前的研究表明,与C31相比,Jcasp在细胞毒性中似乎起次要作用。特别是,用γ-分泌酶抑制剂处理抑制Jcasp的产生,并不会导致C31诱导的毒性有任何减弱。其次,由于在缺乏内源性APP的细胞中基本不存在C31毒性,我们使用分裂β-半乳糖苷酶互补测定法来监测蛋白质-蛋白质相互作用,以确定APP相关复合物的存在。我们的结果表明,APP同聚体和C31/APP异聚体复合物均与细胞死亡相关,这表明C31与APP形成复合物,以招募那些启动与细胞毒性相关信号的相互作用伙伴。