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Information on EC 3.4.21.113 - pestivirus NS3 polyprotein peptidase and Organism(s) Classical swine fever virus and UniProt Accession P19712

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Classical swine fever virus
UNIPROT: P19712 not found.
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The taxonomic range for the selected organisms is: Classical swine fever virus
The enzyme appears in selected viruses and cellular organisms
Reaction Schemes
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Leu is conserved at position P1 for all four cleavage sites. Alanine is found at position P1' of the NS4A-NS4B cleavage site, whereas serine is found at position P1' of the NS3-NS4A, NS4B-NS5A and NS5A-NS5B cleavage sites
Synonyms
nonstructural protein ns3, csfv ns3, wnv ns2b/ns3, flavivirus ns3 protease, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
nonstructural protein 3
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pestivirus NS3 protease
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of peptide bond
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-
-
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CAS REGISTRY NUMBER
COMMENTARY hide
37259-58-8
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SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
non-structural protein 4-5 + H2O
?
show the reaction diagram
in trans-cleavage assays using NS4-5 as a substrate, NS3p acts as a fully functional protease that is able to process the polyprotein
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-
?
additional information
?
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NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
non-structural protein 4-5 + H2O
?
show the reaction diagram
in trans-cleavage assays using NS4-5 as a substrate, NS3p acts as a fully functional protease that is able to process the polyprotein
-
-
?
additional information
?
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
the enzyme is well conserved among the Flaviviridae
malfunction
deletion of Leu1781 results in enzyme inactivation
metabolism
internal cleavage within the NS3 moiety is a common process that further extends the functional repertoires of the multifunctional NS2-3 or NS3 and represents another level of the complex polyprotein processing of Flaviviridae
physiological function
evolution
comparison of the Flaviviridae NS3 proteases
malfunction
metabolism
upon infection of the host cell the viral RNA genome is translated into a polyprotein that is processed by cellular and viral proteases into the mature structural (SP) and nonstructural (NS) proteins. For members of the genus Pestivirus the array in the polyprotein is the following: NH2-Npro (N-terminal autoprotease), C (capsid protein, core), Erns (envelope protein RNase secreted), E1, E2, p7, NS2-3 (NS2 and NS3), NS4A, NS4B, NS5A, NS5B-COOH. The N-terminal autoprotease Npro generates its own C-terminus and thereby the N-terminus of the capsid protein core (C). Further cleavages releasing the structural proteins C, Erns, E1 and E2 as well as p7 are mediated by proteases residing in the endoplasmatic reticulum (ER). The cleavage between NS2 and NS3 is catalyzed by an autoprotease in NS2. The activity of the NS2 protease is temporally regulated by a cellular cofactor leading to significant amounts of uncleaved NS2-3 in pestivirus infected cells. The cleavages downstream of NS3, NS4A, NS4B and NS5A are catalyzed by the serine protease domain of NS3 which requires NS4A as cofactor for full proteolytic activity and is termed NS3-4A protease
physiological function
additional information
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
proteolytic modification
autocatalytic intramolecular cleavages of the enzyme, overview. One cleavable peptide bond occurs between Leu1781 and Met1782, giving rise to a helicase subunit of 55 kDa and, depending on the substrate, a NS2-3 fragment of 78 kDa, or a NS3 protease subunit of 26 kDa. A second intramolecular cleavage is mapped to the Leu1748/Lys1749 peptide bond that yields a proteolytically inactive NS3 fragment. Deletion of either of the cleavage site residues resultsin a loss ofRNAinfectivity, indicating the functional importance of amino acid identity at the respective positions
proteolytic modification
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
purified full-length CSFV NS3 with its NS4A PCS covalently tethered to its N terminus through a flexible linker, X-ray diffraction structure determination and analysis at 2.35 A resolution
purified recombinant soluble CSFV NS4A37NS3 complex, X-ray diffraction structure determination and analysis at 3.05-3.21 A resolution
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
K232A
site-directed mutagenesis, helicase active site mutant
S163A
site-directed mutagenesis, protease active site mutant, proteolytically inactive
V132A
mutation of residue 132 of the NS3 protease domain, the gain of function mutation in NS3 maps to a hydrophobic surface patch which interacts with the NS4A-kink region
additional information
construction of recombinant full-length NS3 (residues 1 to 683) followed by the 8 N-terminal residues of NS4A, the NS3 sequence is preceeded by residues 21 to 57 of the protease cofactor NS4A
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli by nickel affinity chromatography, followed by tag cleavage through TEV protease
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Lamp, B.; Riedel, C.; Wentz, E.; Tortorici, M.A.; Ruemenapf, T.
Autocatalytic cleavage within classical swine fever virus NS3 leads to a functional separation of protease and helicase
J. Virol.
87
11872-11883
2013
Classical swine fever virus (P19712)
Manually annotated by BRENDA team
Riedel, C.; Lamp, B.; Heimann, M.; Koenig, M.; Blome, S.; Moennig, V.; Schuettler, C.; Thiel, H.J.; Ruemenapf, T.
The core protein of classical Swine Fever virus is dispensable for virus propagation in vitro
PLoS Pathog.
8
e1002598
2012
Classical swine fever virus (P19712)
Manually annotated by BRENDA team
Zheng, F.; Lu, G.; Li, L.; Gong, P.; Pan, Z.
Uncoupling of protease trans-cleavage and helicase activities in pestivirus NS3
J. Virol.
91
e01094-17
2017
Classical swine fever virus (Q5U8X5), Classical swine fever virus
Manually annotated by BRENDA team
Dubrau, D.; Tortorici, M.A.; Rey, F.A.; Tautz, N.
A positive-strand RNA virus uses alternative protein-protein interactions within a viral protease/cofactor complex to switch between RNA replication and virion morphogenesis
PLoS Pathog.
13
e1006134
2017
Classical swine fever virus (Q5U8X5)
Manually annotated by BRENDA team