Designed semisynthetic protein inhibitors of Ub/Ubl E1 activating enzymes.
Designed semisynthetic protein inhibitors of Ub/Ubl E1 activating enzymes.
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DOI：
10.1021/ja9088549
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发表时间：
2010-02-17
影响因子：
15
通讯作者：
Tan, Derek S.
中科院分区：
文献类型：
--
作者：
Lu, Xuequan;Olsen, Shaun K.;Capili, Allan D.;Cisar, Justin S.;Lima, Christopher D.;Tan, Derek S.
泛素(Ub)和泛素样修饰物(Ubl)激活酶(E1s)的半合成、基于机理的蛋白质抑制剂已被开发出来，以靶向在蛋白质SUMO化和泛素化过程中Ub/Ubl C末端的腺基化和硫酯化。这些抑制剂是通过内含素介导的表达蛋白连接产生的，使用截短的Ub/Ub1蛋白(相扑残基1-94；Ub残基1-71)与C-端硫酸酯，以及具有C-端腺苷类似物和N-端半胱氨酸残基的合成三肽。SUMO-AMSN(4a)和Ub-AMSN(4b)在第一半反应中含有磺胺基团作为同源化合物Ub/Ub1-AMP腺苷中间体中磷酸基团的非水解性模拟物，这些结构分别以剂量依赖的方式选择性地分别抑制SUMO E1和Ub E1。SUMO-AVSN(5a)和Ub-AVSN(5b)含有一种亲电性的乙烯基磺酰胺，旨在捕获进入的E1半胱氨酸亲核试剂(Uba2 Cys173在SUMO E1中；Uba1 Cys593在Ub E1中)，这些结构分别以半胱氨酸亲核依赖的方式选择性、共价并稳定地与SUMO E1和Ub E1交联。这些抑制剂是探索E1功能中突出的机制问题的有力工具，也可用于研究E1酶的生物学功能。
Semisynthetic, mechanism-based protein inhibitors of ubiquitin (Ub) and ubiquitin-like modifier (Ubl) activating enzymes (E1s) have been developed to target E1-catalyzed adenylation and thioesterification of the Ub/Ubl C-terminus during the processes of protein SUMOylation and ubiquitination. The inhibitors were generated by intein-mediated expressed protein ligation, using a truncated Ub/Ubl protein (SUMO residues 1–94; Ub residues 1–71) with a C-terminal thioester, and synthetic tripeptides having a C-terminal adenosine analogue and an N-terminal cysteine residue. SUMO-AMSN (4a) and Ub-AMSN (4b) contain a sulfamide group as a non-hydrolyzable mimic of the phosphate group in the cognate Ub/Ubl-AMP adenylate intermediate in the first half-reaction, and these constructs selectively inhibit SUMO E1 and Ub E1, respectively, in a dose-dependent manner. SUMO-AVSN (5a) and Ub-AVSN (5b) contain an electrophilic vinyl sulfonamide designed to trap the incoming E1 cysteine nucleophile (Uba2 Cys173 in SUMO E1; Uba1 Cys593 in Ub E1) in the second half-reaction, and these constructs selectively, covalently, and stably crosslink to SUMO E1 and Ub E1, respectively, in a cysteine nucleophile-dependent manner. These inhibitors are powerful tools to probe outstanding mechanistic questions in E1 function and can also be used to study the biological functions of E1 enzymes.
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影响因子：
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