Exploring the biological functional mechanism of the HMGB1/TLR4/MD-2 complex by surface plasmon resonance.
Exploring the biological functional mechanism of the HMGB1/TLR4/MD-2 complex by surface plasmon resonance.
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DOI：
10.1186/s10020-018-0023-8
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发表时间：
2018-05-10
期刊：
影响因子：
--
通讯作者：
Al-Abed Y
中科院分区：
文献类型：
--
作者：
He M;Bianchi ME;Coleman TR;Tracey KJ;Al-Abed Y
高迁移率族蛋白1（HMGB 1）首先被鉴定为非组蛋白染色质结合蛋白，当从坏死细胞或活化的白细胞释放时，其作为促炎细胞因子和损伤相关分子模式分子起作用。HMGB 1由两个结构相似的HMG盒组成，包括促炎（B-box）和抗炎（A-box）结构域。巧合的是，A盒还含有充分表征的抗HMGB 1单克隆抗体“2G 7”的表位，其也在多种体内模型中有效抑制HMGB 1介导的炎症。A-box结构域抑制HMGB 1的炎症活性和2G 7结合A-box结构域后发挥抗炎活性的分子机制一直是一个谜。最近，我们证明：1）TLR 4/MD-2受体是HMGB 1介导的细胞因子产生所必需的; 2）HMGB 1-TLR 4/MD-2相互作用受HMGB 1亚型的氧化还原状态控制。我们使用表面等离子体共振（SPR）研究了HMGB 1亚型（氧化还原态）或HMGB 1片段（A-和B-box）与TLR 4/MD-2复合物的相互作用。我们的结果表明：1）完整的HMGB 1通过A-box结构域以高亲和力但可感知的解离速率结合TLR 4; 2）完整的HMGB 1通过B-box结构域以低亲和力但非常慢的解离速率结合MD-2;和3）单独的HMGB 1 A-box结构域比完整的蛋白质更稳定地结合TLR 4，从而通过阻断HMGB 1与TLR 4/MD-2复合物相互作用来拮抗HMGB 1。这些发现不仅提出了HMGB 1与TLR 4/MD-2在两阶段过程中相互作用的模型，而且还解释了A-box结构域和2G 7如何抑制HMGB 1。本文的在线版本（10.1186/s10020-018-0023-8）包含补充材料，可供授权用户使用。
High Mobility Group Box 1 (HMGB1) was first identified as a nonhistone chromatin-binding protein that functions as a pro-inflammatory cytokine and a Damage-Associated Molecular Pattern molecule when released from necrotic cells or activated leukocytes. HMGB1 consists of two structurally similar HMG boxes that comprise the pro-inflammatory (B-box) and the anti-inflammatory (A-box) domains. Paradoxically, the A-box also contains the epitope for the well-characterized anti-HMGB1 monoclonal antibody “2G7”, which also potently inhibits HMGB1-mediated inflammation in a wide variety of in vivo models. The molecular mechanisms through which the A-box domain inhibits the inflammatory activity of HMGB1 and 2G7 exerts anti-inflammatory activity after binding the A-box domain have been a mystery. Recently, we demonstrated that: 1) the TLR4/MD-2 receptor is required for HMGB1-mediated cytokine production and 2) the HMGB1–TLR4/MD-2 interaction is controlled by the redox state of HMGB1 isoforms. We investigated the interactions of HMGB1 isoforms (redox state) or HMGB1 fragments (A- and B-box) with TLR4/MD-2 complex using Surface Plasmon Resonance (SPR) studies. Our results demonstrate that: 1) intact HMGB1 binds to TLR4 via the A-box domain with high affinity but an appreciable dissociation rate; 2) intact HMGB1 binds to MD-2 via the B-box domain with low affinity but a very slow dissociation rate; and 3) HMGB1 A-box domain alone binds to TLR4 more stably than the intact protein and thereby antagonizes HMGB1 by blocking HMGB1 from interacting with the TLR4/MD-2 complex. These findings not only suggest a model whereby HMGB1 interacts with TLR4/MD-2 in a two-stage process but also explain how the A-box domain and 2G7 inhibit HMGB1. The online version of this article (10.1186/s10020-018-0023-8) contains supplementary material, which is available to authorized users.
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影响因子：
10.6
作者：
Kang, Rui;Chen, Ruochan;Zhang, Qiuhong;Hou, Wen;Wu, Sha;Cao, Lizhi;Huang, Jin;Yu, Yan;Fan, Xue-gong;Yan, Zhengwen;Sun, Xiaofang;Wang, Haichao;Wang, Qingde;Tsung, Allan;Billiar, Timothy R.;Zeh, Herbert J., III;Lotze, Michael T.;Tang, Daolin
通讯作者：
Tang, Daolin
影响因子：
3.5
作者：
Sahu, Debashish;Debnath, Priyanka;Takayama, Yuki;Iwahara, Junji
通讯作者：
Iwahara, Junji
影响因子：
2.2
作者：
Li, JH;Wang, HC;Yang, H
通讯作者：
Yang, H
影响因子：
2.8
作者：
Visintin, Alberto;Iliev, Dimitar B.;Golenbock, Douglas T.
通讯作者：
Golenbock, Douglas T.
DOI：
10.1016/j.bbrc.2013.10.085
发表时间：
2013-11-29
影响因子：
3.1
作者：
Wang, Jing;Tochio, Naoya;Tate, Shin-ichi
通讯作者：
Tate, Shin-ichi