Detection of ubiquitin-proteasome enzymatic activities in cells: application of activity-based probes to inhibitor development.
Detection of ubiquitin-proteasome enzymatic activities in cells: application of activity-based probes to inhibitor development.
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DOI：
10.1016/j.bbamcr.2012.05.014
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发表时间：
2012-11
期刊：
影响因子：
--
通讯作者：
Altun M
中科院分区：
文献类型：
--
作者：
Kramer HB;Nicholson B;Kessler BM;Altun M
背景：模拟天然底物的合成探针能够检测细胞环境中的酶活性。这种基于活性的探针已被应用的一个领域是泛素-蛋白酶体途径，其正在成为重要的治疗靶点。已经开发了一系列试剂，其特异性地标记去泛素化酶（DUB）并促进其抑制剂的表征。审查范围：在这里，我们专注于应用探针的细胞内DUBs，一组特定的蛋白酶参与泛素蛋白酶体系统。特别是，这个家族的蛋白酶，特异性地识别泛素和泛素样蛋白的活性亚基的功能表征将进行讨论。此外，我们提出了潜在的和设计的基于活性的探针靶向激酶和磷酸酶研究磷酸化。主要结论：合成分子探针增加了我们对DUBs在活细胞中的功能作用的理解。除了检测已知成员的酶活性外，基于活性的探针还有助于以前未表征的酶的许多功能分配。该方法能够对小分子DUB抑制剂的特异性进行细胞验证。一般意义：分子探针结合基于质谱的蛋白质组学和细胞测定代表了发现和功能验证的强大方法，这一概念可以扩展到其他酶类。这解决了对加速药物开发过程所需的更多信息的基于细胞的测定的需求。这篇文章是一个特殊问题的一部分，题为：泛素药物发现和诊断。基于活性的化学探针筛选细胞中小分子去泛素化酶抑制剂的特异性。通过质谱法对去泛素化酶特异性抑制剂进行基于细胞的分析。蛋白酶体、去泛素化酶、激酶和磷酸酶探针概述。
Background: Synthetic probes that mimic natural substrates can enable the detection of enzymatic activities in a cellular environment. One area where such activity-based probes have been applied is the ubiquitin–proteasome pathway, which is emerging as an important therapeutic target. A family of reagents has been developed that specifically label deubiquitylating enzymes (DUBs) and facilitate characterization of their inhibitors. Scope of review: Here we focus on the application of probes for intracellular DUBs, a group of specific proteases involved in the ubiquitin proteasome system. In particular, the functional characterization of the active subunits of this family of proteases that specifically recognize ubiquitin and ubiquitin-like proteins will be discussed. In addition we present the potential and design of activity-based probes targeting kinases and phosphatases to study phosphorylation. Major conclusions: Synthetic molecular probes have increased our understanding of the functional role of DUBs in living cells. In addition to the detection of enzymatic activities of known members, activity-based probes have contributed to a number of functional assignments of previously uncharacterized enzymes. This method enables cellular validation of the specificity of small molecule DUB inhibitors. General significance: Molecular probes combined with mass spectrometry-based proteomics and cellular assays represent a powerful approach for discovery and functional validation, a concept that can be expanded to other enzyme classes. This addresses a need for more informative cell-based assays that are required to accelerate the drug development process. This article is part of a Special Issue entitled: Ubiquitin Drug Discovery and Diagnostics. ► Activity-based chemical probe screen for specificity of small molecule deubiquitylating enzyme inhibitors in cells. ► Cell-based profiling of inhibitors specific for deubiquitylating enzymes by mass spectrometry. ► Overview of proteasome, deubiquitylating enzyme, kinase, and phosphatase probes.
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影响因子：
3.6
作者：
Artavanis-Tsakonas, Katerina;Misaghi, Shahram;Ploegh, Hidde L.
通讯作者：
Ploegh, Hidde L.
影响因子：
--
作者：
Greenbaum, D;Medzihradszky, KF;Bogyo, M
通讯作者：
Bogyo, M
影响因子：
--
作者：
Bogyo, M;Shin, S;Ploegh, HL
通讯作者：
Ploegh, HL
影响因子：
--
作者：
Greenbaum, DC;Arnold, WD;Bogyo, M
通讯作者：
Bogyo, M
DOI：
10.1084/jem.187.1.97
发表时间：
1998-01-05
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Griffin TA;Nandi D;Cruz M;Fehling HJ;Kaer LV;Monaco JJ;Colbert RA
通讯作者：
Colbert RA