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Dengue virus 3 NS5 methyltransferase domain: expression, purification, crystallization and first structural data from microcrystalline specimens

  • Alexandros Valmas , Stavroula Fili , Nikos Nikolopoulos , Maria Spiliopoulou , Magdalini Christopoulou , Fotini Karavassili , Christos Kosinas , Konstantinos Bastalias , Eleftheria Rosmaraki , Julie Lichiére , Andrew Fitch , Detlef Beckers , Thomas Degen , Nicolas Papageorgiou , Bruno Canard , Bruno Coutard und Irene Margiolaki EMAIL logo
Veröffentlicht/Copyright: 2. November 2017
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Zeitschrift für Kristallographie - Crystalline Materials
Aus der Zeitschrift Zeitschrift für Kristallographie - Crystalline Materials Band 233 Heft 5

Abstract

Flavivirus infections often provoke life-threatening diseases of epidemic magnitudes, thus extensive research is currently directed towards the development of efficient vaccines and approved antiviral compounds. We present here the expression, purification, crystallization and preliminary X-ray diffraction analysis of one of the components of the flavivirus replication complex, the non-structural protein 5 (NS5) mRNA methyltransferase (MTase) domain, from an emerging pathogenic flavivirus, dengue virus 3 (DEN3). Polycrystalline precipitates of DEN3 NS5 MTase, suitable for X-ray powder diffraction (XRPD) measurements, were produced in the presence of PEG 8000 (25–32.5% (w/v)), 0.1 M Tris-Amino, in a pH range from 7.0 to 8.0. A polymorph of orthorhombic symmetry (space group: P21212, a=61.9 Å, b=189.6 Å, c=52.4 Å) was identified via XRPD. These results are the first step towards the complete structural determination of this molecule via XRPD and a parallel demonstration of the applicability of the method.

Keywords: dengue virus 3; methyltransferase; non structural protein 5; powder diffraction; structural virology

Acknowledgements

We would like to thank the ESRF for provision of beamtime at the ID22 beamline and Dr. Jonathan P. Wright (ID11) for his multiannual scientific collaboration. We would like also to thank PANalytical for instrumentation and software support and, the European Virus Archive goes Global (EVAg) (European Union H2020 Grant Agreement No 653316) for providing the expression plasmid. This research has been financially supported by General Secretariat for Research and Technology (GSRT) and the Hellenic Foundation for Research and Innovation (HFRI) (Scholarship Code: 2467) and co-financed by the following grants: the European Union (European Social Fund) in collaboration with Greek State, under the ‘ARISTEIA II’ Action (MIS Code 4659) of the ‘Operational Program Education And Lifelong Learning’, the European Union (European Regional Development Fund – ERDF) and Greek national funds through the Operational Program ‘Regional Operational Program’ of the National Strategic Reference Framework (NSRF), the International Atomic Energy Agency (CRP code F12024) and the COST Action (CM1306). Finally, the company NanoMEGAS supported this work.

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Received: 2017-7-25
Accepted: 2017-9-25
Published Online: 2017-11-2
Published in Print: 2018-5-24

©2018 Walter de Gruyter GmbH, Berlin/Boston

Artikel in diesem Heft

  1. Frontmatter
  2. Graphical Synopsis
  3. Inorganic Crystal Structures
  4. The ternary phases Cu1.5−xLi1+x Sb and Cu4+xLi1−x Sb2, and their structural relations
  5. Synthesis and crystal structures of the new ternary borides Fe3Al2B2 and Ru9Al3B8 and the confirmation of Ru4Al3B2 and Ru9Al5B8−x (x≈2)
  6. Dengue virus 3 NS5 methyltransferase domain: expression, purification, crystallization and first structural data from microcrystalline specimens
  7. The truth is out there: the metal-π interactions in crystal of Cr(CO)3(pcp) as revealed by the study of vibrational smearing of electron density
  8. Role of defectivity on the crystallography of martensitic transformations in Ti50Ni40Cu10: an XRD investigation
  9. Organic and Metalorganic Crystal Structures
  10. Structure–property relation and third-order nonlinear optical studies of two new halogenated chalcones
https://doi.org/10.1515/zkri-2017-2091
Schlagwörter für diesen Artikel
dengue virus 3; methyltransferase; non structural protein 5; powder diffraction; structural virology

Artikel in diesem Heft

  1. Frontmatter
  2. Graphical Synopsis
  3. Inorganic Crystal Structures
  4. The ternary phases Cu1.5−xLi1+x Sb and Cu4+xLi1−x Sb2, and their structural relations
  5. Synthesis and crystal structures of the new ternary borides Fe3Al2B2 and Ru9Al3B8 and the confirmation of Ru4Al3B2 and Ru9Al5B8−x (x≈2)
  6. Dengue virus 3 NS5 methyltransferase domain: expression, purification, crystallization and first structural data from microcrystalline specimens
  7. The truth is out there: the metal-π interactions in crystal of Cr(CO)3(pcp) as revealed by the study of vibrational smearing of electron density
  8. Role of defectivity on the crystallography of martensitic transformations in Ti50Ni40Cu10: an XRD investigation
  9. Organic and Metalorganic Crystal Structures
  10. Structure–property relation and third-order nonlinear optical studies of two new halogenated chalcones
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