Self-inhibited State of Venezuelan Equine Encephalitis Virus (VEEV) nsP2 Cysteine Protease: A Crystallographic and Molecular Dynamics Analysis.
Self-inhibited State of Venezuelan Equine Encephalitis Virus (VEEV) nsP2 Cysteine Protease: A Crystallographic and Molecular Dynamics Analysis.
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DOI：
10.1016/j.jmb.2023.168012
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发表时间：
2023-03-15
影响因子：
5.6
通讯作者：
Motyan, Janos Andras
中科院分区：
文献类型：
--
作者：
Hoffka, Gyula;Lountos, George T.;Needle, Danielle;Wlodawer, Alexander;Waugh, David S.;Tozser, Jozsef;Motyan, Janos Andras
委内瑞拉马脑炎病毒(VEEV)属于Togaviridae科，对人和马都有致病性。VEEV非结构蛋白2(NsP2)是一种处理多聚蛋白的半胱氨酸蛋白酶(NsP2pro)，因此它是发现抑制剂的药物靶点。先前通过X射线结晶学和计算研究表征了VEEV nsP2催化域的原子结构。在N端含有N475A突变的修饰的nsP2pro显示出一种意想不到的构象：N末端残基与活性部位结合，类似于底物的结合。由于N475对N末端和活性部位的稳定起着重要作用，推测N475A突变可能导致N末端的较大构象变化。这种构象首先在N475A突变体中观察到，但我们在确定含有野生型N475活性中心残基和K741A/K767A表面熵减少突变的催化活性nsP2pro的晶体结构时也发现了这种构象。这表明N475A突变不是自我抑制的先决条件。在这里，我们描述了一个截短的nsP2pro(残基463-785，K741A/K767A)的高分辨(1.46？)晶体结构，并进一步用分子动力学分析了nsP2pro及其N475A突变体的活性构象和自抑制构象。N-末端残基的不同构象的比较有助于了解在酶的稳定中起重要作用的相互作用。
The Venezuelan equine encephalitis virus (VEEV) belongs to the Togaviridae family and is pathogenic to both humans and equines. The VEEV non-structural protein 2 (nsP2) is a cysteine protease (nsP2pro) that processes the polyprotein and thus it is a drug target for inhibitor discovery. The atomic structure of the VEEV nsP2 catalytic domain was previously characterized by both X-ray crystallography and computational studies. A modified nsP2pro harboring a N475A mutation in the N terminus was observed to exhibit an unexpected conformation: the N-terminal residues bind to the active site, mimicking binding of a substrate. The large conformational change of the N terminus was assumed to be induced by the N475A mutation, as N475 has an important role in stabilization of the N terminus and the active site. This conformation was first observed in the N475A mutant, but we also found it while determining a crystal structure of the catalytically active nsP2pro containing the wild-type N475 active site residue and K741A/K767A surface entropy reduction mutations. This suggests that the N475A mutation is not a pre-requisite for self-inhibition. Here, we describe a high resolution (1.46 Å) crystal structure of a truncated nsP2pro (residues 463–785, K741A/K767A) and analyze the structure further by molecular dynamics to study the active and self-inhibited conformations of nsP2pro and its N475A mutant. A comparison of the different conformations of the N-terminal residues sheds a light on the interactions that play an important role in the stabilization of the enzyme.
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影响因子：
5.6
作者：
Bozóki B;Mótyán JA;Hoffka G;Waugh DS;Tőzsér J
通讯作者：
Tőzsér J
影响因子：
4.6
作者：
Ahmed A;Bakovic A;Risner K;Kortchak S;Der Torossian Torres M;de la Fuente-Nunez C;Lu T;Bhalla N;Narayanan A
通讯作者：
Narayanan A
影响因子：
4.4
作者：
JORGENSEN, WL;CHANDRASEKHAR, J;KLEIN, ML
通讯作者：
KLEIN, ML
影响因子：
5.4
作者：
Balistreri, Giuseppe;Caldentey, Javier;Ahola, Tero
通讯作者：
Ahola, Tero
影响因子：
6.3
作者：
Le Grand, Scott;Goetz, Andreas W.;Walker, Ross C.
通讯作者：
Walker, Ross C.