Literature summary for 3.4.21.91 extracted from

Junaid, M.; Angsuthanasombat, C.; Wikberg, J.E.; Ali, N.; Katzenmeier, G.
A straightforward experimental approach to expression, purification, refolding, and enzymatic analysis of recombinant dengue virus NS2B(H)-NS3pro protease (2013), Biochemistry (Moscow), 78, 920-924.

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Cloned(Commentary)

Cloned (Comment)	Organism
expression of NS2B(H)-NS3pro complex in E.scherichiacoli	Dengue virus

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
0.1	-	acetyl-Nle-Lys-Arg-Arg-4-methylcoumarin 7-amide	pH 9.5, 37°C	Dengue virus

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
10000	-	1 * 21000 plus 1 * 10000, SDS-PAGE, corresponding to the NS3pro protease domain only, and the His-tagged NS2B(H) cofactor	Dengue virus
21000	-	1 * 21000 plus 1 * 10000, SDS-PAGE, corresponding to the NS3pro protease domain only, and the His-tagged NS2B(H) cofactor	Dengue virus
37000	-	x * 37000, SDS-PAGE, intact NS2B(H)-NS3pro complex	Dengue virus

Organism

Organism	UniProt	Comment	Textmining
Dengue virus	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
expression of NS2B(H)-NS3pro complex in E.scherichiacoli predominantly as insoluble protein and purification to 95% purity	Dengue virus

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
acetyl-Nle-Lys-Arg-Arg-4-methylcoumarin 7-amide + H2O	-	Dengue virus	acetyl-Nle-Lys-Arg-Arg + 7-amino-4-methylcoumarin	-	?

Subunits

Subunits	Comment	Organism
?	x * 37000, SDS-PAGE, intact NS2B(H)-NS3pro complex	Dengue virus
dimer	1 * 21000 plus 1 * 10000, SDS-PAGE, corresponding to the NS3pro protease domain only, and the His-tagged NS2B(H) cofactor	Dengue virus

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
0.112	-	acetyl-Nle-Lys-Arg-Arg-4-methylcoumarin 7-amide	pH 9.5, 37°C	Dengue virus

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism	Structure
1.12	-	acetyl-Nle-Lys-Arg-Arg-4-methylcoumarin 7-amide	pH 9.5, 37°C	Dengue virus

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Release 2023.1 (January 2023)