Characterization of a caspase-3-substrate kinome using an N- and C-terminally tagged protein kinase library produced by a cell-free system.
Characterization of a caspase-3-substrate kinome using an N- and C-terminally tagged protein kinase library produced by a cell-free system.
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DOI：
10.1038/cddis.2010.65
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发表时间：
2010-10-28
影响因子：
9
通讯作者：
中科院分区：
文献类型：
--
作者：
胱天蛋白酶-3（CASP 3）切割许多蛋白质，包括蛋白激酶（PK）。了解CASP 3及其PK底物之间的关系对于描述由CASP 3活性控制的凋亡信号级联是必要的。我们在本文中报告了使用简单的无细胞系统合成包含在其N-和C-末端标记的304个PK（NC标记的PK）的文库和报告CASP 3切割事件的发光测定的CASP 3底物激酶组的表征。发现43个PK（包括30个新鉴定的PK）是CASP 3底物，并确定了23个PK中的28个切割位点。有趣的是，23个PK中有16个在其N-或C-末端的60个残基内具有切割位点。此外，29个PKs在凋亡细胞中裂解，包括5个在体外靠近其末端裂解。总的来说，大约14%的PK测试是CASP 3底物，这表明PK的CASP 3裂解可能是一个标志性事件在抗肿瘤信号级联。该蛋白水解测定方法将鉴定其他蛋白酶底物。
Caspase-3 (CASP3) cleaves many proteins including protein kinases (PKs). Understanding the relationship(s) between CASP3 and its PK substrates is necessary to delineate the apoptosis signaling cascades that are controlled by CASP3 activity. We report herein the characterization of a CASP3-substrate kinome using a simple cell-free system to synthesize a library that contained 304 PKs tagged at their N- and C-termini (NCtagged PKs) and a luminescence assay to report CASP3 cleavage events. Forty-three PKs, including 30 newly identified PKs, were found to be CASP3 substrates, and 28 cleavage sites in 23 PKs were determined. Interestingly, 16 out of the 23 PKs have cleavage sites within 60 residues of their N- or C-termini. Furthermore, 29 of the PKs were cleaved in apoptotic cells, including five that were cleaved near their termini in vitro. In total, approximately 14% of the PKs tested were CASP3 substrates, suggesting that CASP3 cleavage of PKs may be a signature event in apoptotic-signaling cascades. This proteolytic assay method would identify other protease substrates.
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影响因子：
64.5
作者：
Mahrus S;Trinidad JC;Barkan DT;Sali A;Burlingame AL;Wells JA
通讯作者：
Wells JA
影响因子：
4.4
作者：
Matsuoka, Kazuhiro;Komori, Hiroaki;Nose, Masato;Endo, Yaeta;Sawasaki, Tatsuya
通讯作者：
Sawasaki, Tatsuya
影响因子：
11.4
作者：
Emoto, Y;Manome, Y;Kufe, D
通讯作者：
Kufe, D
DOI：
10.1016/j.bbrc.2009.12.175
发表时间：
2010-02-12
影响因子：
3.1
作者：
Ohoka, Nobumichi;Sakai, Satoshi;Hayashi, Hidetoshi
通讯作者：
Hayashi, Hidetoshi
影响因子：
64.5
作者：
Dix MM;Simon GM;Cravatt BF
通讯作者：
Cravatt BF