Proteome-wide identification of family memberspecific natural substrate repertoire of caspases
Proteome-wide identification of family memberspecific natural substrate repertoire of caspases
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DOI：
10.1073/pnas.0702251104
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发表时间：
2007-09-04
影响因子：
11.1
通讯作者：
Liu, Rihe
中科院分区：
文献类型：
--
作者：
Ju, Wujian;Valencia, C. Alexander;Liu, Rihe
半胱天冬酶是细胞凋亡所必需的蛋白水解酶。了解许多离散和相互作用的信号通路介导的半胱天冬酶需要识别的天然底物库为每个半胱天冬酶的利益。使用一种基于扩增的蛋白质选择技术，称为mRNA展示，我们开发了一个高通量筛选平台，在蛋白质组范围内的caspase家族成员特异性底物。从人类蛋白质组中鉴定出大量已知的和以前未表征的半胱天冬酶-3底物。这些选定的底物，包括其切割位点和特异性的蛋白水解功能，其特征在于。容易选择仅被胱天蛋白酶-8或颗粒酶B裂解而不被胱天蛋白酶-3裂解的底物。该方法可以广泛应用于有效和系统地鉴定感兴趣的生物体中的任何半胱天冬酶的家族成员特异性天然底物库，以及具有高特异性的许多其他蛋白酶的家族成员特异性天然底物库。
Caspases are proteolytic enzymes that are essential for apoptosis. Understanding the many discrete and interacting signaling pathways mediated by caspases requires the identification of the natural substrate repertoire for each caspase of interest. Using an amplification-based protein selection technique called mRNA display, we developed a high-throughput screen platform for caspase family member specific substrates on a proteome-wide scale. A large number of both known and previously uncharacterized caspase-3 substrates were identified from the human proteome. The proteolytic features of these selected substrates, including their cleavage sites and specificities, were characterized. Substrates that were cleaved only by caspase-8 or granzyme B but not by caspase-3, were readily selected. The method can be widely applied for efficient and systematic identification of the family member specific natural substrate repertoire of any caspase in an organism of interest, in addition to that of numerous other proteases with high specificity.