The dengue virus NS2B-NS3 protease retains the closed conformation in the complex with BPTI

FEBS Lett. 2014 Jun 27;588(14):2206-11. doi: 10.1016/j.febslet.2014.05.018. Epub 2014 May 21.

Abstract

The C-terminal β-hairpin of NS2B (NS2Bc) in the dengue virus NS2B-NS3 protease is required for full enzymatic activity. In crystal structures without inhibitor and in the complex with bovine pancreatic trypsin inhibitor (BPTI), NS2Bc is displaced from the active site. In contrast, nuclear magnetic resonance (NMR) studies in solution only ever showed NS2Bc in the enzymatically active closed conformation. Here we demonstrate by pseudocontact shifts from a lanthanide tag that NS2Bc remains in the closed conformation also in the complex with BPTI. Therefore, the closed conformation is the best template for drug discovery.

Keywords: Bovine pancreatic trypsin inhibitor; Dengue virus protease; Lanthanide tag; NMR spectroscopy; Pseudocontact shift.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aprotinin / chemistry*
  • Catalytic Domain
  • Dengue Virus / enzymology*
  • Models, Molecular
  • Nuclear Magnetic Resonance, Biomolecular
  • Protein Structure, Secondary
  • RNA Helicases / chemistry
  • Serine Endopeptidases / chemistry
  • Viral Nonstructural Proteins / chemistry*

Substances

  • NS2B protein, flavivirus
  • NS3 protein, flavivirus
  • Viral Nonstructural Proteins
  • Aprotinin
  • Serine Endopeptidases
  • RNA Helicases