Literature summary for 3.6.4.13 extracted from

Luo, D.; Xu, T.; Watson, R.P.; Scherer-Becker, D.; Sampath, A.; Jahnke, W.; Yeong, S.S.; Wang, C.H.; Lim, S.P.; Strongin, A.; Vasudevan, S.G.; Lescar, J.
Insights into RNA unwinding and ATP hydrolysis by the flavivirus NS3 protein (2008), EMBO J., 27, 3209-3219.

Activating Compound

Activating Compound	Comment	Organism	Structure
single-stranded DNA	on single-stranded RNA binding, the NS3 enzyme switches to a catalytic competent state imparted by an inward movement of the P-loop, interdomain closure and a change in the divalent metal coordination shell, providing a structural basis for RNA-stimulated ATP hydrolysis	Dengue virus

Cloned(Commentary)

Cloned (Comment)	Organism
expression in Escherichia coli	Dengue virus
expression of the His-tagged catalytic domain of the NS3 helicase domain from dengue virus serotype 4 in Escherichia coli strain BL21	Dengue virus

Crystallization (Commentary)

Crystallization (Comment)	Organism
hanging drop vapour diffusion method, crystallization of native enzyme, enzyme in complex with adenylyl imidodiphosphate, enzyme in complex with ADP, enzyme in complex with single-stranded RNA and enzyme in complex with single-stranded RNA and ADP	Dengue virus
purified recombinant His-tagged catalytic domain of the NS3, hanging drop vapour diffusion method, at 13°C over a well solution containing 0.1M MES, pH 6.5, and 20% PEG 3350, X-ray diffraction structure determination and analysis. Crystals for the AMPPNP complex are obtained by cocrystallization of NS3h at 5 mg/ml with 5 mM MnCl2 and 5 mM AMPPNP using a precipitating solution containing 0.1M MES, pH 6.5, and 10% PEG 3350, at 13°C. Crystals with ADP are obtained by cocrystallization at a concentration of 2.5 mg/ml with 5 mM MnCl2 and 5 mM ADP in 0.1 M Tris-HCl, pH 7.0, and 7.5% PEG 3350 at 23°C, further preparation of ternary complexes,overview	Dengue virus

Crystallization (Comment)

Organism

hanging drop vapour diffusion method, crystallization of native enzyme, enzyme in complex with adenylyl imidodiphosphate, enzyme in complex with ADP, enzyme in complex with single-stranded RNA and enzyme in complex with single-stranded RNA and ADP

Dengue virus

purified recombinant His-tagged catalytic domain of the NS3, hanging drop vapour diffusion method, at 13°C over a well solution containing 0.1M MES, pH 6.5, and 20% PEG 3350, X-ray diffraction structure determination and analysis. Crystals for the AMPPNP complex are obtained by cocrystallization of NS3h at 5 mg/ml with 5 mM MnCl2 and 5 mM AMPPNP using a precipitating solution containing 0.1M MES, pH 6.5, and 10% PEG 3350, at 13°C. Crystals with ADP are obtained by cocrystallization at a concentration of 2.5 mg/ml with 5 mM MnCl2 and 5 mM ADP in 0.1 M Tris-HCl, pH 7.0, and 7.5% PEG 3350 at 23°C, further preparation of ternary complexes,overview

Dengue virus

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mg2+	activates, binding complexes, overview	Dengue virus

Organism

Organism	UniProt	Comment	Textmining
Dengue virus	-	-	-
Dengue virus	-	serotype 4	-

Purification (Commentary)

Purification (Comment)	Organism
-	Dengue virus
recombinant His-tagged catalytic domain of the NS3 helicase domain from dengue virus serotype 4 from Escherichia coli strain BL21 by nickel affinity chromatography and gel filtration	Dengue virus

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
ATP + H2O	-	Dengue virus	ADP + phosphate	-	?
additional information	conformational changes during ATP hydrolysis and RNA unwinding: on ssRNA binding, the NS3 enzyme switches to a catalytic competent state imparted by an inward movement of the P-loop, interdomain closure and a change in the divalent metal coordination shell, providing a structural basis for RNA-stimulated ATP hydrolysis. Determination of enzyme structure-function relationship of enzyme bound to single-stranded RNA, to an ATP analogue, to a transition-state analogue and to ATP hydrolysis products. RNA recognition appears largely sequence independent, reaction mechanism and RNA recognition, overview. RNA-unwinding mechanism, overview	Dengue virus	?	-	?

Synonyms

Synonyms	Comment	Organism
NS3 helicase	-	Dengue virus
NS3 protein	-	Dengue virus