Chemistry-based functional proteomics reveals novel members of the deubiquitinating enzyme
Chemistry-based functional proteomics reveals novel members of the deubiquitinating enzyme
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DOI：
10.1016/s1074-5521(02)00248-x
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发表时间：
2002-10-01
影响因子：
--
通讯作者：
Kessler, BM
中科院分区：
文献类型：
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作者：
Borodovsky, A;Ovaa, H;Kessler, BM
泛素-蛋白酶体系统包括一个大家族的去泛素化酶（DUBs）。许多成员被分配到这类酶的序列相似性，但没有生物活性的证据。一组新的dub特异性探针通过化学结扎法产生。这些探针可以通过串联质谱法鉴定dub和相关成分，并快速证明从初级结构推断功能的基因产物的酶活性。我们在EL4细胞中发现了23个活性dub，包括肿瘤抑制因子CYLD1。至少有两个dub与蛋白酶体19S调节复合体紧密相互作用。分离到一个OTU结构域蛋白，与已知dub序列无同源性。我们发现这种多肽与Ub的C端发生反应，从而证明了这种新型蛋白酶超家族的类似dub的酶活性。
The ubiquitin (Ub)-proteasome system includes a large family of deubiquitinating enzymes (DUBs). Many members are assigned to this enzyme class by sequence similarity but without evidence for biological activity. A panel of novel DUB-specific probes was generated by a chemical ligation method. These probes allowed identification of DUBs and associated components by tandem mass spectrometry, as well as rapid demonstration of enzymatic activity for gene products whose functions were inferred from primary structure. We identified 23 active DUBs in EL4 cells, including the tumor suppressor CYLD1. At least two DUBs tightly interact with the proteasome 19S regulatory complex. An OTU domain-containing protein, with no sequence homology to any known DUBs, was isolated. We show that this polypeptide reacts with the C terminus of Ub, thus demonstrating DUB-like enzymatic activity for this novel superfamily of proteases.