Small ubiquitin-related modifier (SUMO)-specific proteases -: Profiling the specificities and activities of human SENPs
Small ubiquitin-related modifier (SUMO)-specific proteases -: Profiling the specificities and activities of human SENPs
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DOI：
10.1074/jbc.m702444200
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发表时间：
2007-09-07
影响因子：
4.8
通讯作者：
Salvesen, Guy S.
中科院分区：
文献类型：
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作者：
Mikolajczyk, Jowita;Drag, Marcin;Salvesen, Guy S.
SENP是通过产生成熟的小泛素相关修饰物（SUMO）用于蛋白质缀合（内肽酶活性）和从靶标去除缀合的SUMO（异肽酶活性）来参与SUMO化调节的蛋白酶。使用纯化的重组催化结构域的7个人SENP中的6个，我们证明了它们各自的活性对SUMO-1，-2和-3的特异性。识别底物的主要模式是通过SUMO结构域，C-末端尾引导内肽酶特异性。一般来说，SENP 1是最有效的内肽酶，而SENP 2和-5-7具有比内肽酶活性高得多的异肽酶活性。我们开发了荧光四肽底物，SENP裂解，使我们能够表征每种酶的环境概况。使用这些合成底物，我们揭示了SUMO结构域增强SENP 1，-2，-5，-6和-7的催化，证明SUMO对SENP的底物诱导活化。
SENPs are proteases that participate in the regulation of SUMOylation by generating mature small ubiquitin-related modifiers ( SUMO) for protein conjugation ( endopeptidase activity) and removing conjugated SUMO from targets ( isopeptidase activity). Using purified recombinant catalytic domains of 6 of the 7 human SENPs, we demonstrate the specificity of their respective activities on SUMO-1, -2, and -3. The primary mode of recognition of substrates is via the SUMO domain, and the C-terminal tails direct endopeptidase specificity. Broadly speaking, SENP1 is the most efficient endopeptidase, whereas SENP2 and -5-7 have substantially higher isopeptidase than endopeptidase activities. We developed fluorogenic tetrapeptide substrates that are cleaved by SENPs, enabling us to characterize the environmental profiles of each enzyme. Using these synthetic substrates we reveal that the SUMO domain enhances catalysis of SENP1, -2, -5,-6, and -7, demonstrating substrate-induced activation of SENPs by SUMOs.