Selective Inhibition of Cysteine-Dependent Enzymes by Bioorthogonal Tethering.
Selective Inhibition of Cysteine-Dependent Enzymes by Bioorthogonal Tethering.
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DOI：
10.1016/j.jmb.2022.167524
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发表时间：
2022-03
影响因子：
5.6
通讯作者：
Luke A. Spear;Yang Huang;Jinghao Chen-;Alexander R Nödling;S. Virdee;Y. Tsai
中科院分区：
文献类型：
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作者：
Luke A. Spear;Yang Huang;Jinghao Chen-;Alexander R Nödling;S. Virdee;Y. Tsai
利用一种常见的工具化合物快速和选择性地抑制细胞中的酶的一般方法将对研究和治疗开发具有重要价值。我们之前报道了一种化学发生策略，该策略解决了激酶的这一挑战，依赖于通过遗传编码的非规范氨基酸将泛素抑制剂与靶激酶的生物正交系结。然而，泛酶抑制剂对许多酶类是不可用的。在这里，我们通过亲电弹头的生物正交系绳将化学发生策略的范围扩展到半胱氨酸依赖性酶。为了证明这一概念，在生化分析中证明了两种E2泛素结合酶UBE2L3和UBE2D1的选择性抑制作用。该方法的进一步发展和优化将使其能够在细胞中以及与其他半胱氨酸依赖酶一起使用，促进其细胞功能的研究和作为治疗靶点的验证。
A general approach for the rapid and selective inhibition of enzymes in cells using a common tool compound would be of great value for research and therapeutic development. We previously reported a chemogenetic strategy that addresses this challenge for kinases, relying on bioorthogonal tethering of a pan inhibitor to a target kinase through a genetically encoded non-canonical amino acid. However, pan inhibitors are not available for many enzyme classes. Here, we expand the scope of the chemogenetic strategy to cysteine-dependent enzymes by bioorthogonal tethering of electrophilic warheads. For proof of concept, selective inhibition of two E2 ubiquitin-conjugating enzymes, UBE2L3 and UBE2D1, was demonstrated in biochemical assays. Further development and optimization of this approach should enable its use in cells as well as with other cysteine-dependent enzymes, facilitating the investigation of their cellular function and validation as therapeutic targets.