Synthesis and Application of Activity-Based Probes for Proteases.

Synthesis and Application of Activity-Based Probes for Proteases.
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蛋白酶活性探针的合成及应用

DOI:
10.1007/978-1-4939-6850-3_19
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发表时间:
2017
影响因子:
--
通讯作者:
S. H. L.
S. H. L.
中科院分区:
--
文献类型:
--
作者:
Van Kersavond;Nguyen;Verhelst;S. H. L.

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活性蛋白酶的检测、可视化和鉴定可以通过基于活性的探针来促进,所述探针共价结合靶蛋白酶的催化残基。基于活性的探针的合成可能具有挑战性。在这里,我们概述了一个简单的协议,探针合成的基础上,标准的固相肽合成,然后用反应性亲电试剂作为弹头的N-末端的上限。通过用Western印迹或荧光扫描作为读数标记细胞和组织裂解物中的半胱氨酸蛋白酶来说明探针的适用性。
The detection, visualization, and identification of active proteases can be facilitated by activity-based probes, which covalently bind to a catalytic residue of the target protease. The synthesis of activity-based probes can be challenging. We here outline a simple protocol for probe synthesis based on standard solid phase peptide synthesis followed by capping of the N-terminus with a reactive electrophile as a warhead. The applicability of the probes is illustrated by labeling cysteine proteases in cell and tissue lysates with Western blotting or fluorescence scanning as a readout.
DOI: 10.1074/mcp.m112.021014
发表时间: 2013-01-01
影响因子: 7
作者:
Yang, Yinliang;Hahne, Hannes;Verhelst, Steven H. L.
通讯作者: Verhelst, Steven H. L.
DOI: 10.1016/s1074-5521(00)00014-4
发表时间: 2000-08-01
影响因子: --
作者:
Greenbaum, D;Medzihradszky, KF;Bogyo, M
通讯作者: Bogyo, M
细胞裂解物中点击化学介导的基于活性的蛋白质分析的比较分析
DOI: --
发表时间: 2013
期刊: Molecules
影响因子: 4.6
作者:
Yinliang Yang;Xiaomeng Yang;S. Verhelst
通讯作者: S. Verhelst