柯萨奇病毒B3蛋白酶3C：表达、纯化、结晶及初步结构解析
Coxsackievirus B3 protease 3C: expression, purification, crystallization and preliminary structural insights.
作者信息
Fili Stavroula, Valmas Alexandros, Christopoulou Magdalini, Spiliopoulou Maria, Nikolopoulos Nikos, Lichière Julie, Logotheti Souzana, Karavassili Fotini, Rosmaraki Eleftheria, Fitch Andrew, Wright Jonathan, Beckers Detlef, Degen Thomas, Nénert Gwilherm, Hilgenfeld Rolf, Papageorgiou Nicolas, Canard Bruno, Coutard Bruno, Margiolaki Irene
机构信息
Section of Genetics, Cell Biology and Development, Department of Biology, University of Patras, University Campus, 26500 Patras, Greece.
Architecture et Fonction des Macromolécules Biologiques, CNRS and Universités d'Aix-Marseille I et II, UMR 6098, ESIL Case 925, 13288 Marseille, France.
出版信息
Acta Crystallogr F Struct Biol Commun. 2016 Dec 1;72(Pt 12):877-884. doi: 10.1107/S2053230X16018513. Epub 2016 Nov 25.
Viral proteases are proteolytic enzymes that orchestrate the assembly of viral components during the viral life cycle and proliferation. Here, the expression, purification, crystallization and preliminary X-ray diffraction analysis are presented of protease 3C, the main protease of an emerging enterovirus, coxsackievirus B3, that is responsible for many cases of viral myocarditis. Polycrystalline protein precipitates suitable for X-ray powder diffraction (XRPD) measurements were produced in the presence of 22-28%(w/v) PEG 4000, 0.1 M Tris-HCl, 0.2 M MgCl in a pH range from 7.0 to 8.5. A polymorph of monoclinic symmetry (space group C2, unit-cell parameters a = 77.9, b = 65.7, c = 40.6 Å, β = 115.9°) was identified via XRPD. These results are the first step towards the complete structural determination of the molecule via XRPD and a parallel demonstration of the accuracy of the method.
病毒蛋白酶是在病毒生命周期和增殖过程中协调病毒组件组装的蛋白水解酶。本文介绍了一种新出现的肠道病毒——柯萨奇病毒B3的主要蛋白酶3C的表达、纯化、结晶及初步X射线衍射分析，该病毒导致了许多病毒性心肌炎病例。在22 - 28%（w/v）聚乙二醇4000、0.1 M Tris - HCl、0.2 M MgCl₂存在的条件下，于pH值7.0至8.5范围内产生了适用于X射线粉末衍射（XRPD）测量的多晶蛋白质沉淀。通过XRPD鉴定出一种单斜对称的多晶型物（空间群C2，晶胞参数a = 77.9，b = 65.7，c = 40.6 Å，β = 115.9°）。这些结果是通过XRPD对该分子进行完整结构测定的第一步，也是该方法准确性的平行证明。
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