title: The dengue virus NS2B-NS3 protease retains the closed conformation in the complex with BPTI.
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.
authors.affiliation: Australian National University, Research School of Chemistry, Canberra, ACT 0200, Australia.
authors.forename: Wan-Na
authors.initials: WN
authors.lastname: Chen
authors.affiliation: Australian National University, Research School of Chemistry, Canberra, ACT 0200, Australia.
authors.forename: Karin V
authors.initials: KV
authors.lastname: Loscha
authors.affiliation: Medicinal Chemistry, Institute of Pharmacy and Molecular Biotechnology, Heidelberg University, Im Neuenheimer Feld 364, 69120 Heidelberg, Germany.
authors.forename: Christoph
authors.initials: C
authors.lastname: Nitsche
authors.affiliation: Medicinal Chemistry and Drug Action, Monash Institute of Pharmaceutical Sciences, Parkville, VIC 3052, Australia.
authors.forename: Bim
authors.initials: B
authors.lastname: Graham
authors.affiliation: Australian National University, Research School of Chemistry, Canberra, ACT 0200, Australia. Electronic address: gottfried.otting@anu.edu.au.
authors.forename: Gottfried
authors.initials: G
authors.lastname: Otting
mesh_terms: D007611:Aprotinin / Q000737:chemistry*; D020134:Catalytic Domain; D003716:Dengue Virus / Q000201:enzymology*; D008958:Models, Molecular; D019906:Nuclear Magnetic Resonance, Biomolecular; D017433:Protein Structure, Secondary; D020365:RNA Helicases / Q000737:chemistry; D012697:Serine Endopeptidases / Q000737:chemistry; D017361:Viral Nonstructural Proteins / Q000737:chemistry*; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases
chemical_list: D007611:Aprotinin; D020365:RNA Helicases; D012697:Serine Endopeptidases; D017361:Viral Nonstructural Proteins; C080102:NS2B protein, flavivirus; C060028:NS3 protein, flavivirus; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases
keywords: Bovine pancreatic trypsin inhibitor; Dengue virus protease; Lanthanide tag; NMR spectroscopy; Pseudocontact shift