TY - JOUR
T1 - Coxsackievirus B3 protease 3C: Expression, purification, crystallization and preliminary structural insights
AU - Fili, Stavroula
AU - Valmas, Alexandros
AU - Christopoulou, Magdalini
AU - Spiliopoulou, Maria
AU - Nikolopoulos, Nikos
AU - Lichière, Julie
AU - Logotheti, Souzana
AU - Karavassili, Fotini
AU - Rosmaraki, Eleftheria
AU - Fitch, Andrew
AU - Wright, Jonathan
AU - Beckers, Detlef
AU - Degen, Thomas
AU - Nénert, Gwilherm
AU - Hilgenfeld, Rolf
AU - Papageorgiou, Nicolas
AU - Canard, Bruno
AU - Coutard, Bruno
AU - Margiolaki, Irene
PY - 2016/12/1
Y1 - 2016/12/1
N2 - 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 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.The expression, purification, crystallization, X-ray powder diffraction data collection and preliminary analysis of protease 3C from coxsackievirus B3 are reported.
AB - 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 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.The expression, purification, crystallization, X-ray powder diffraction data collection and preliminary analysis of protease 3C from coxsackievirus B3 are reported.
UR - http://www.scopus.com/inward/record.url?scp=85002427149&partnerID=8YFLogxK
U2 - 10.1107/S2053230X16018513
DO - 10.1107/S2053230X16018513
M3 - Journal articles
C2 - 27917835
AN - SCOPUS:85002427149
SN - 2053-230X
VL - 72
SP - 877
EP - 884
JO - Acta Crystallographica Section:F Structural Biology Communications
JF - Acta Crystallographica Section:F Structural Biology Communications
IS - 12
ER -
