Conformational dynamics linked to domain closure and substrate binding explain the ERAP1 allosteric regulation mechanism.
Conformational dynamics linked to domain closure and substrate binding explain the ERAP1 allosteric regulation mechanism.
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DOI：
10.1038/s41467-021-25564-w
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发表时间：
2021-09-06
影响因子：
16.6
通讯作者：
Stern LJ
中科院分区：
文献类型：
--
作者：
Maben Z;Arya R;Georgiadis D;Stratikos E;Stern LJ
内质网氨基肽酶ERAP1处理抗原肽，以装载在MHC-I蛋白上，并被CD8 T细胞识别，因为它们检查身体的感染和恶性肿瘤。晶体结构揭示了ERAP1的开放或闭合构象，但这些结构是否存在于溶液中并参与催化尚不清楚。在这里，我们通过小角X射线散射来评估ERAP1在底物、变构激活剂和抑制剂存在的溶液中的构象状态。我们还用X射线结晶学表征了蛋白质构象的变化，并通过化学交联法定位了替代的C末端结合位点。结构和酶数据表明，ERAP1活性位点的结构重构在物理上与结构域的关闭有关，并通过与长肽底物的结合而促进。这些结果阐明了ERAP1催化所需的步骤，证明了构象动力学在催化循环中的重要性，并为观察到的变构调节和Lys/Arg528多态疾病关联提供了机制。内质网氨基肽酶ERAP1处理多肽，用于抗原呈递。在这里，作者评估了ERAP1在溶液中的构象状态，提供了对ERAP1底物长度依赖的催化活性和调节的分子机制的见解，包括自身免疫性疾病相关多态性的影响。
The endoplasmic-reticulum aminopeptidase ERAP1 processes antigenic peptides for loading on MHC-I proteins and recognition by CD8 T cells as they survey the body for infection and malignancy. Crystal structures have revealed ERAP1 in either open or closed conformations, but whether these occur in solution and are involved in catalysis is not clear. Here, we assess ERAP1 conformational states in solution in the presence of substrates, allosteric activators, and inhibitors by small-angle X-ray scattering. We also characterize changes in protein conformation by X-ray crystallography, and we localize alternate C-terminal binding sites by chemical crosslinking. Structural and enzymatic data suggest that the structural reconfigurations of ERAP1 active site are physically linked to domain closure and are promoted by binding of long peptide substrates. These results clarify steps required for ERAP1 catalysis, demonstrate the importance of conformational dynamics within the catalytic cycle, and provide a mechanism for the observed allosteric regulation and Lys/Arg528 polymorphism disease association. The endoplasmic-reticulum aminopeptidase ERAP1 processes peptides for antigen presentation. Here, the authors assess ERAP1 conformational states in solution, providing insight into the molecular mechanisms of ERAP1 substrate-length dependent catalytic activity and regulation, including the effects of autoimmune disease-associated polymorphism.
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DOI：
10.1073/pnas.1210123109
发表时间：
2012-10-30
影响因子：
11.1
作者：
Chen, Lang;Lin, Yi-Lun;Li, Fang
通讯作者：
Li, Fang
影响因子：
4.7
作者：
Chen, Bin;Li, Dahe;Xu, Weidong
通讯作者：
Xu, Weidong
影响因子：
3.5
作者：
Goto, Y;Hattori, A;Tsujimoto, M
通讯作者：
Tsujimoto, M
DOI：
10.1107/s0907444909052925
发表时间：
2010-02
期刊：
Acta crystallographica. Section D, Biological crystallography
影响因子：
--
作者：
Adams PD;Afonine PV;Bunkóczi G;Chen VB;Davis IW;Echols N;Headd JJ;Hung LW;Kapral GJ;Grosse-Kunstleve RW;McCoy AJ;Moriarty NW;Oeffner R;Read RJ;Richardson DC;Richardson JS;Terwilliger TC;Zwart PH
通讯作者：
Zwart PH
影响因子：
5.5
作者：
Bettencourt, Bruno Filipe;Rocha, Fabiana Leal;Bruges-Armas, Jacome
通讯作者：
Bruges-Armas, Jacome