Coxsackievirus B3 protease 3C: expression, purification, crystallization and preliminary structural insights. Issue 12 (1st December 2016)

• Record Type: Journal Article
• Authors: 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
• Is Part Of: Acta crystallographica. Volume 72:Issue 12(2016:Dec.)
• Page Start: 877
• Page End: 884
• Publication Date: 2016-12-01
• Subjects: 3C protease -- coxsackievirus B3 -- powder diffraction; Crystallography -- Periodicals; Crystals -- Periodicals; 548
• DOI: 10.1107/S2053230X16018513
• Languages: English
• ISSNs: 2053-230X

Abstract

Abstract : The expression, purification, crystallization, X‐ray powder diffraction data collection and preliminary analysis of protease 3C from coxsackievirus B3 are reported. Abstract : 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 MgCl2 in a pH range from 7.0 to 8.5. A polymorph of monoclinic symmetry (space group C 2, 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.