Literature summary for 3.4.21.91 extracted from

de la Cruz, L.; Nguyen, T.H.; Ozawa, K.; Shin, J.; Graham, B.; Huber, T.; Otting, G.
Binding of Low Molecular Weight Inhibitors Promotes Large Conformational Changes in the Dengue Virus NS2B-NS3 Protease: Fold Analysis by Pseudocontact Shifts (2011), J. Am. Chem. Soc., 133, 19205-19215.

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Cloned(Commentary)

Cloned (Comment)	Organism
the dengue 2 NS2B-NS3pro construct contains 255 residues, including a (Gly)4-Ser-(Gly)4 linker between NS2B (47 residues) and NS3pro (185 residues). In addition, it contains a His-tag at the C-terminus and the T7 gene 10 N-terminal peptide MASMTG at the N-terminus followed by a two-residue cloning artifact (LE), resulting in a 27 kDa protein. To suppress slow autocleavage detected by SDS-PAGE, most samples have the mutation Lys15Ala which cause no significant changes in the appearance of the NMR spectra. Protein is expressed in Escherichia coli	Dengue virus type 2

Cloned (Comment)

Organism

the dengue 2 NS2B-NS3pro construct contains 255 residues, including a (Gly)4-Ser-(Gly)4 linker between NS2B (47 residues) and NS3pro (185 residues). In addition, it contains a His-tag at the C-terminus and the T7 gene 10 N-terminal peptide MASMTG at the N-terminus followed by a two-residue cloning artifact (LE), resulting in a 27 kDa protein. To suppress slow autocleavage detected by SDS-PAGE, most samples have the mutation Lys15Ala which cause no significant changes in the appearance of the NMR spectra. Protein is expressed in Escherichia coli

Dengue virus type 2

Protein Variants

Protein Variants	Comment	Organism
K117R/T122R	mutant is analyzed by 15N-HSQC spectra with and without inhibitor 4-guanidino-benzoic acid-4-nitrophenyl ester	Dengue virus type 2
K15A	autocleavage detected by SDS-PAGE is supressed	Dengue virus type 2
additional information	16 possible mutants bearing and an exchanged Ile to Val are analyzed by 15N-HSQC spectra with and without inhibitor 4-guanidino-benzoic acid-4-nitrophenyl ester	Dengue virus type 2
additional information	7 Ser to Ala mutants are analyzed by 15N-HSQC spectra with and without inhibitor 4-guanidino-benzoic acid-4-nitrophenyl ester	Dengue virus type 2

Inhibitors

Inhibitors	Comment	Organism
4-guanidino-benzoic acid 4-nitrophenyl ester	the fold of Dengue NS2B-NS3pro in solution with and without bound inhibitor by nuclear magnetic resonance spectroscopy is analyzed. Multiple paramagnetic lanthanide tags are attached to different sites to generate pseudocontact shifts (PCS). The PCSs show, that in the presence of a positively charged low-molecular weight inhibitor, the enzyme assumes a closed state that is very similar to the closed state previously observed for the West Nile virus protease. To assess the open state, a binding site for a Gd3+ complex is created and paramagnetic relaxation enhancements is measured. The results show that the specific open conformation displayed in the crystal of DEN NS2B-NS3pro is barely populated in solution	Dengue virus type 2
5-methyl-2-[4-[(4-[[4-(4-methyl-4,5-dihydro-1H-imidazol-3-ium-2-yl)phenyl]amino]-4a,8a-dihydrophthalazin-1-yl)amino]phenyl]-4,5-dihydro-1H-imidazol-3-ium	-	Dengue virus type 2
benzoyl-Nle-Lys-Arg-Arg	-	Dengue virus type 2

Organism

Organism	UniProt	Comment	Textmining
Dengue virus type 2	-	-	-

Renatured (Commentary)

Renatured (Comment)	Organism
using Ni-NTA spin columns	Dengue virus type 2

Synonyms

Synonyms	Comment	Organism
NS2B-NS3 protease	-	Dengue virus type 2