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Coxsackievirus B3 protease 3C: expression, purification, crystallization and preliminary structural insights
Author(s) -
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
Publication year - 2016
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 37
ISSN - 2053-230X
DOI - 10.1107/s2053230x16018513
Subject(s) - protease , crystallization , proteases , monoclinic crystal system , crystallography , chemistry , powder diffraction , materials science , enzyme , biochemistry , crystal structure , organic chemistry
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 2 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.