Allosteric Inhibitors, Crystallography, and Comparative Analysis Reveal Network of Coordinated Movement across Human Herpesvirus Proteases.
Allosteric Inhibitors, Crystallography, and Comparative Analysis Reveal Network of Coordinated Movement across Human Herpesvirus Proteases.
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DOI：
10.1021/jacs.7b04030
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发表时间：
2017-08-30
影响因子：
15
通讯作者：
Craik CS
中科院分区：
文献类型：
--
作者：
Acker TM;Gable JE;Bohn MF;Jaishankar P;Thompson MC;Fraser JS;Renslo AR;Craik CS
靶向隐结合位点是一种有吸引力但尚未开发的小分子调节蛋白质功能的方法。利用卡波西肉瘤相关疱疹病毒（KSHV）的二聚体蛋白酶（Pr）作为模型系统，我们试图剖析一个假定的变弹性网络，将二聚界面的一个隐位点与酶功能连接起来。利用变构抑制剂与二聚体界面位点结合，得到了5个低温x射线结构。在室温x射线数据和其他二聚体疱疹病毒蛋白酶的比较分析中，变构抑制剂捕获的不同协调运动也显示为替代状态。通过详细的动力学分析阐明了两步机制，并提出了一种酶异构化抑制模型。最后，从这类的一个代表性的变构抑制剂被证明是有效的细胞模型的病毒感染。这些研究揭示了一个连接KHSV Pr的隐结合位点占用和变构失活的原子通信协调动态网络，可以用来靶向该临床相关酶家族的其他成员。
Targeting of cryptic binding sites represents an attractive but underexplored approach to modulating protein function with small molecules. Using the dimeric protease (Pr) from Kaposi’s sarcoma-associated herpes-virus (KSHV) as a model system, we sought to dissect a putative allosteric network linking a cryptic site at the dimerization interface to enzyme function. Five cryogenic X-ray structures were solved of the monomeric protease with allosteric inhibitors bound to the dimer interface site. Distinct coordinated movements captured by the allosteric inhibitors were also revealed as alternative states in room-temperature X-ray data and comparative analyses of other dimeric herpesvirus proteases. A two-step mechanism was elucidated through detailed kinetic analyses and suggests an enzyme isomerization model of inhibition. Finally, a representative allosteric inhibitor from this class was shown to be efficacious in a cellular model of viral infectivity. These studies reveal a coordinated dynamic network of atomic communication linking cryptic binding site occupancy and allosteric inactivation of KHSV Pr that can be exploited to target other members of this clinically relevant family of enzymes.
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影响因子：
6.7
作者：
Zühlsdorf M;Werten S;Klupp BG;Palm GJ;Mettenleiter TC;Hinrichs W
通讯作者：
Hinrichs W
影响因子：
5.6
作者：
Lee, Gregory M.;Shahian, Tina;Craik, Charles S.
通讯作者：
Craik, Charles S.
影响因子：
62.1
作者：
Gable, Jonathan E.;Acker, Timothy M.;Craik, Charles S.
通讯作者：
Craik, Charles S.
DOI：
10.1073/pnas.1111325108
发表时间：
2011-09-27
影响因子：
11.1
作者：
Fraser, James S.;van den Bedem, Henry;Alber, Tom
通讯作者：
Alber, Tom
影响因子：
5.4
作者：
PRESTON, VG;COATES, JAV;RIXON, FJ
通讯作者：
RIXON, FJ