Recognition of Lys48-Linked Di-ubiquitin and Deubiquitinating Activities of the SARS Coronavirus Papain-like Protease.
Recognition of Lys48-Linked Di-ubiquitin and Deubiquitinating Activities of the SARS Coronavirus Papain-like Protease.
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DOI：
10.1016/j.molcel.2016.04.016
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发表时间：
2016-05-19
期刊：
影响因子：
16
通讯作者：
Lima CD
中科院分区：
文献类型：
--
作者：
Békés M;van der Heden van Noort GJ;Ekkebus R;Ovaa H;Huang TT;Lima CD
去泛素化酶（DUB）识别和切割连接特异性聚泛素（polyUb）链，但在许多情况下，特异性的机制仍然难以捉摸。严重急性呼吸综合征（SARS）冠状病毒木瓜蛋白酶样蛋白酶（PLpro）是一种DUB，可切割ISG 15（一种双结构域的Ub样蛋白）和Lys 48连接的polyUb链，释放diUbLys 48产物。为了阐明这种特异性，我们报告了SARS PLpro与基于diUbLys 48活性的探针结合的2.85 kDa晶体结构。SARS PLpro通过两个接触位点S1和S2以延伸构象结合diUbLys 48，S1和S2分别位于活性位点的近端和远端。我们发现，polyUbLys 48链的特异性是基于S2位点的接触，并通过S1-S1′优先选择跨越活性位点的Lys 48连接来增强。相比之下，ISG 15特异性由S1位点中的接触主导。polyUbLys 48特异性的决定因素应该被证明有助于理解PLpro在冠状病毒感染中的去泛素化活性。一种基于Lys 48连接特异性双泛素活性的探针选择性标记SARS PLpro双UbLys 48与SARS PLpro复合物的结构揭示了一种扩展的双Ub构象S2-S1和S1-S1′相互作用使SARS PLpro对K48连接的多聚泛素具有特异性SARS PLpro通过不同的方式识别Lys 48连接的polyUb链和ISG 15 Békés et al.提出了SARS病毒PLpro与diUbLys 48复合物的高分辨率晶体结构，揭示了Lys 48连接的diUb单元的延伸构象，并显示了SARS PLpro对Lys 48连接的polyUb链的偏好的生物化学基础。
Deubiquitinating enzymes (DUBs) recognize and cleave linkage-specific polyubiquitin (polyUb) chains, but mechanisms underlying specificity remain elusive in many cases. The severe acute respiratory syndrome (SARS) coronavirus papain-like protease (PLpro) is a DUB that cleaves ISG15, a two-domain Ub-like protein, and Lys48-linked polyUb chains, releasing diUbLys48 products. To elucidate this specificity, we report the 2.85 Å crystal structure of SARS PLpro bound to a diUbLys48 activity-based probe. SARS PLpro binds diUbLys48 in an extended conformation via two contact sites, S1 and S2, which are proximal and distal to the active site, respectively. We show that specificity for polyUbLys48 chains is predicated on contacts in the S2 site and enhanced by an S1-S1′ preference for a Lys48 linkage across the active site. In contrast, ISG15 specificity is dominated by contacts in the S1 site. Determinants revealed for polyUbLys48 specificity should prove useful in understanding PLpro deubiquitinating activities in coronavirus infections. A Lys48 linkage-specific diubiquitin activity-based probe selectively labels SARS PLpro The structure of a diUbLys48∼SARS PLpro complex reveals an extended di-Ub conformation S2-S1 and S1-S1′ interactions make SARS PLpro specific for K48-linked polyubiquitin SARS PLpro recognizes Lys48-linked polyUb chains and ISG15 via distinct manners Békés et al. present a high-resolution crystal structure of a SARS virus PLpro∼diUbLys48 complex that reveals an extended conformation of the Lys48-linked diUb unit and shows the biochemical basis for SARS PLpro’s preference for Lys48-linked polyUb chains.