Literature summary for 3.4.21.91 extracted from

Lorenz, I.C.; Marcotrigiano, J.; Dentzer, T.G.; Rice, C.M.
Structure of the catalytic domain of the hepatitis C virus NS2-3 protease (2006), Nature, 442, 831-835.

Search for more literature for 3.4.21.91

Application

Application	Comment	Organism
drug development	proteolysis through formation of a composite active site in NS2-3 protease occurs in the context of the viral polyprotein expressed in mammalian cells. This offers new opportunities for antiviral drug design	Hepacivirus C

Cloned(Commentary)

Cloned (Comment)	Organism
NS2pro (NS2 residues 94-217), a truncated form of NS2 shown to be proteolytically active in the context of an NS2-3 precursor that includes the NS3 protease domain, expressed in Escherichia coli. NS2 containing wild-type or mutant active site residues expressed in mammalian cells in the context of a full-length viral polyprotein or as NS2-3 precursor with an N-terminal Flag or HA tag	Hepacivirus C

Crystallization (Commentary)

Crystallization (Comment)	Organism
by hanging-drop vapour diffusion at 4°C, at 2.3 A resolution. Crystal structure of the catalytic domain of NS2-3 protease, using native and selenomethionine-containing protein yields two crystal forms with the same space group P21. Dimeric cysteine protease with two composite active sites. For each active site, the catalytic histidine and glutamate residues are contributed by one monomer, and the nucleophilic cysteine by the other. Carboxy-terminal residues remain coordinated in the two active sites, predicting an inactive post-cleavage form	Hepacivirus C

Crystallization (Comment)

Organism

by hanging-drop vapour diffusion at 4°C, at 2.3 A resolution. Crystal structure of the catalytic domain of NS2-3 protease, using native and selenomethionine-containing protein yields two crystal forms with the same space group P21. Dimeric cysteine protease with two composite active sites. For each active site, the catalytic histidine and glutamate residues are contributed by one monomer, and the nucleophilic cysteine by the other. Carboxy-terminal residues remain coordinated in the two active sites, predicting an inactive post-cleavage form

Hepacivirus C

Protein Variants

Protein Variants	Comment	Organism
C184A	mutation in the NS2 active site, is defective in NS2-3 processing. When Hepatitis C virus polyproteins with NS2 containing a C184A mutation are co-expressed, NS2 and NS3 cleavage products are detected	Hepacivirus C
H143A	mutation in the NS2 active site, is defective in NS2-3 processing. When Hepatitis C virus polyproteins with NS2 containing a H143A mutation are co-expressed, NS2 and NS3 cleavage products are detected	Hepacivirus C
H143A/C184A	mixing wild-type Flag-NS2-3 with double-mutant HA-NS2-3(H143A/C184A) or vice versa yields only cleaved wild-type NS2, whereas the double-mutant polypeptide remains unprocessed because neither of the composite active sites is functional, when a wild-type and a double-mutant NS2-3 dimerize	Hepacivirus C

Organism

Organism	UniProt	Comment	Textmining
Hepacivirus C	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
by immobilized-metal affinity chromatography, ion exchange chromatography and gel filtration	Hepacivirus C

Subunits

Subunits	Comment	Organism
dimer	crystallography	Hepacivirus C

Synonyms

Synonyms	Comment	Organism
NS2-3 protease	-	Hepacivirus C