Genetically Encoded Crosslinking Enables Identification of Multivalent Ubiquitin-Deubiquitylating Enzyme Interactions.
Genetically Encoded Crosslinking Enables Identification of Multivalent Ubiquitin-Deubiquitylating Enzyme Interactions.
复制标题
基因编码交联能够鉴定多价泛素-去泛素化酶相互作用。
DOI：
10.1002/cbic.202300305
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发表时间：
2023
期刊：
影响因子：
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通讯作者：
Das,Chittaranjan
中科院分区：
文献类型：
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作者：
Patel,Rishi;NegrónTerón,Kristos;Zhou,Mowei;Nakayasu,Ernesto;Drown,Bryon;Das,Chittaranjan
泛素（Ub）蛋白质型通过其多样性控制真核细胞生物学的几乎每个方面。受广泛使用的Ub C末端亲电体（Ub−E）的启发，我们在这里报告了使用遗传密码扩展（GCE）和交联质谱法鉴定Ub与去泛素化酶（Dubs）的多价结合。虽然Ub−Es仅收集S1 Dub位点的结构信息，但我们证明Ub与对苯甲酰基-L-苯丙氨酸的GCE能够识别S1位点以外的相互作用模式，并具有真核和原核来源的Dubs。总的来说，这代表了下一代基于Ub的亲和探针，具有独特的能力，能够揭示基于半胱氨酸的化学物质所提供的Ub相互作用。
Ubiquitin (Ub) proteoforms control nearly every aspect of eukaryotic cell biology through their diversity. Inspired by the widely used Ub C‐terminal electrophiles (Ub−E), here we report the identification of multivalent binding of Ub with deubiquitylating enzymes (Dubs) using genetic code expansion (GCE) and crosslinking mass spectrometry. While the Ub−Es only gather structural information with the S1 Dub sites, we demonstrate that GCE of Ub with p‐benzoyl‐L‐phenylalanine enables identification of interaction modes beyond the S1 site with a panel of Dubs of both eukaryotic and prokaryotic origin. Collectively, this represents the next generation of Ub‐based affinity probes with a unique ability to unravel Ub interaction landscapes beyond what is afforded by cysteine‐based chemistries.