Molecular basis of ubiquitin-specific protease 8 autoinhibition by the WW-like domain.
Molecular basis of ubiquitin-specific protease 8 autoinhibition by the WW-like domain.
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DOI：
10.1038/s42003-021-02802-x
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发表时间：
2021-11-08
影响因子：
5.9
通讯作者：
Fukushima T
中科院分区：
文献类型：
--
作者：
Kakihara K;Asamizu K;Moritsugu K;Kubo M;Kitaguchi T;Endo A;Kidera A;Ikeguchi M;Kato A;Komada M;Fukushima T
泛素特异性蛋白水解酶8(USP8)是一种脱泛素酶，参与多种膜转运途径。该酶的活性受14-3-3蛋白结合的抑制。USP8中14-3-3结合基序的突变与库欣病有关。然而，USP8活性调控的分子基础仍不清楚。根据本研究中进行的下拉和单分子FRET分析的结果，本研究确定USP8的645-684氨基酸是一个可能与催化USP结构域相互作用的自抑制区域。在计算机模拟中，该区域形成了一种类似WW的结构域结构，堵塞了催化裂隙，并缩小了泛素结合口袋的入口。此外，14-3-3抑制USP8活性的部分原因是增强了WW样结构域和USP结构域之间的相互作用。这些发现为USP8通过类WW结构域自抑制提供了分子基础。此外，他们认为，由于USP8突变，自身抑制的释放可能是库欣病的基础。为了加深我们对USP8调控的理解，Kakihara等人鉴定了USP8的645-684氨基酸，并对其进行了鉴定，USP8是一个自身抑制区。他们的下拉和单分子FRET分析，以及在计算机模拟中，表明USP8自身抑制的释放可能是库欣病的基础。
Ubiquitin-specific protease 8 (USP8) is a deubiquitinating enzyme involved in multiple membrane trafficking pathways. The enzyme activity is inhibited by binding to 14-3-3 proteins. Mutations in the 14-3-3-binding motif in USP8 are related to Cushing’s disease. However, the molecular basis of USP8 activity regulation remains unclear. This study identified amino acids 645–684 of USP8 as an autoinhibitory region, which might interact with the catalytic USP domain, as per the results of pull-down and single-molecule FRET assays performed in this study. In silico modelling indicated that the region forms a WW-like domain structure, plugs the catalytic cleft, and narrows the entrance to the ubiquitin-binding pocket. Furthermore, 14-3-3 inhibited USP8 activity partly by enhancing the interaction between the WW-like and USP domains. These findings provide the molecular basis of USP8 autoinhibition via the WW-like domain. Moreover, they suggest that the release of autoinhibition may underlie Cushing’s disease due to USP8 mutations. In order to advance our understanding of the regulation of Ubiquitin-specific protease 8 (USP8), which is known to play a role in Cushing’s Disease, Kakihara et al identify and characterise amino acids 645–684 of USP8, which serve as an autoinhibitory region. Their pull-down and single-molecule FRET analysis, as well as in silico modelling, suggest that the release of USP8 autoinhibition may underlie Cushing’s disease.
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