Structure of prothrombin in the closed form reveals new details on the mechanism of activation.
Structure of prothrombin in the closed form reveals new details on the mechanism of activation.
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DOI：
10.1038/s41598-018-21304-1
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发表时间：
2018-02-13
影响因子：
4.6
通讯作者：
Pozzi N
中科院分区：
文献类型：
--
作者：
Chinnaraj M;Chen Z;Pelc LA;Grese Z;Bystranowska D;Di Cera E;Pozzi N
凝血因子凝血酶原存在于封闭和开放构象之间的平衡，但这些形式的生理作用仍不清楚。至于其他变构蛋白，分子转换和功能之间的联系的阐明是由试剂稳定在每一个替代的构象。凝血酶原的开放形式在结构上已经被表征，但是关于在生理条件下在溶液中占主导地位的封闭形式的结构知之甚少。利用X射线晶体学和单分子FRET，我们表征了通过工程二硫键锁定在闭合构象中的凝血酶原构建体。结构：（i）提供了最近报道的Kringle-1在蛋白酶结构域上的分子内塌陷的结构验证;（ii）记录了连接Kringle-1和Kringle-2的接头在稳定封闭形式中的关键作用;和（iii）揭示了将平衡向开放构象转移的新机制。与功能研究一起，我们的研究结果定义了封闭和开放构象在凝血酶原转化为凝血酶中的作用，并建立了凝血酶原激活的分子框架，使与凝血酶原突变相关的现有表型合理化，并指出了新的治疗干预策略。
The clotting factor prothrombin exists in equilibrium between closed and open conformations, but the physiological role of these forms remains unclear. As for other allosteric proteins, elucidation of the linkage between molecular transitions and function is facilitated by reagents stabilized in each of the alternative conformations. The open form of prothrombin has been characterized structurally, but little is known about the architecture of the closed form that predominates in solution under physiological conditions. Using X-ray crystallography and single-molecule FRET, we characterize a prothrombin construct locked in the closed conformation through an engineered disulfide bond. The construct: (i) provides structural validation of the intramolecular collapse of kringle-1 onto the protease domain reported recently; (ii) documents the critical role of the linker connecting kringle-1 to kringle-2 in stabilizing the closed form; and (iii) reveals novel mechanisms to shift the equilibrium toward the open conformation. Together with functional studies, our findings define the role of closed and open conformations in the conversion of prothrombin to thrombin and establish a molecular framework for prothrombin activation that rationalizes existing phenotypes associated with prothrombin mutations and points to new strategies for therapeutic intervention.
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影响因子：
7.5
作者：
Morrissey JH;Davis-Harrison RL;Tavoosi N;Ke K;Pureza V;Boettcher JM;Clay MC;Rienstra CM;Ohkubo YZ;Pogorelov TV;Tajkhorshid E
通讯作者：
Tajkhorshid E
凝血酶原向凝血酶的过渡。
DOI：
10.1111/jth.12217
发表时间：
2013-06
期刊：
Journal of thrombosis and haemostasis : JTH
影响因子：
--
作者：
Krishnaswamy S
通讯作者：
Krishnaswamy S
DOI：
10.1107/s0907444904019158
发表时间：
2004-12-01
影响因子：
2.2
作者：
Emsley, P;Cowtan, K
通讯作者：
Cowtan, K
影响因子：
10.4
作者：
Jayandharan, G;Viswabandya, A;Srivastava, A
通讯作者：
Srivastava, A
影响因子：
4.8
作者：
Kroh, Heather K.;Panizzi, Peter;Bock, Paul E.
通讯作者：
Bock, Paul E.