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Information on EC 3.4.21.115 - infectious pancreatic necrosis birnavirus Vp4 peptidase and Organism(s) Infectious pancreatic necrosis virus and UniProt Accession Q703G9

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This record set is specific for:
Infectious pancreatic necrosis virus
UNIPROT: Q703G9 not found.
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The taxonomic range for the selected organisms is: Infectious pancreatic necrosis virus
The enzyme appears in selected viruses and cellular organisms
Reaction Schemes
cleaves the (Ser/Thr)-Xaa-Ala-/-(Ser/Ala)-Gly motif in the polyprotein NH2-pVP2-VP4-VP3-COOH of infectious pancreatic necrosis virus at the pVP2-VP4 and VP4-VP3 junctions
Synonyms
ns protease, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
infectious pancreatic necrosis birnavirus Vp4 protease
-
equine arterivirus serine peptidase
-
-
-
-
infectious pancreatic necrosis birnavirus Vp4 protease
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of peptide bond
-
hydrolysis of peptide bond
-
-
CAS REGISTRY NUMBER
COMMENTARY hide
330588-95-9
-
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
polyprotein of IPNV + H2O
pVP2 + VP3 + VP4
show the reaction diagram
cleaves at the (Ser/Thr)-X-Ala-/-(Ser/Ala)-Gly motif, -/- = cleavage site
-
?
polyprotein of BSNV + H2O
pVP2 + VP3 + VP4
show the reaction diagram
-
-
-
-
?
polyprotein of IPNV + H2O
pVP2 + VP3 + VP4
show the reaction diagram
-
the viral NS protease is active and cleaves the polyprotein when it is expressed in Escherichia coli
-
-
?
additional information
?
-
acyl-enzyme complex for intermolecular self-cleavage reaction determined by crystallization, conserved overall structure of VP4 determined despite of only 20% sequence identity between different VP4 proteins
-
-
?
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
polyprotein of BSNV + H2O
pVP2 + VP3 + VP4
show the reaction diagram
-
-
-
-
?
polyprotein of IPNV + H2O
pVP2 + VP3 + VP4
show the reaction diagram
-
the viral NS protease is active and cleaves the polyprotein when it is expressed in Escherichia coli
-
-
?
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
first identification of an acyl-enzyme complex for a Ser/Lys dyad protease, catalytic mechanism and substrate recognition indicated
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
20500
recombinant protein VP4_524-716-K674A (VP4hex), 194 residues long, theoretical isoelectric point of 4.3 estimated
21700
recombinant protein VP4_514-716-K674A (VP4tri), 204 residues long, theoretical isoelectric point of4.5 estimated
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
proteolytic modification
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
two crystal structures of the IPNV VP4 protease solved from two different crystal symmetries, refinement to 2.2 A resolution reveals the acyl-enzyme complex formed with an internal VP4 cleavage site, substrate binding pockets S1, S3, S5, and S6 identified, conserved overall structure of VP4 determined by three-dimensional alignments
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D537Q
mutant with wild-type activity
D585I
mutant with wild-type activity
D595L
mutant with wild-type activity
D601S
mutant with wild-type activity
D644I
mutant with wild-type activity
D660G/D661S
mutant with wild-type activity
D672N
mutant with wild-type activity
D693L
reduced activity for the VP4-VP3 junction
H547S
reduced activity for the VP4-VP3 junction
H697L
reduced activity for the VP4-VP3 junction
H704S
mutant with wild-type activity
K674A
two recombinant constructs VP4_524-716-K674A and VP4_514-716-K674A generated by site-directed mutagenesis
S633C
40% reduction of VP2-VP4 cleavage, 80% reduction of VP4-VP3 cleavage
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
gel filtration, native and recombinant proteins
eluted from SDS-PAGE gel
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli Tuner (DE3) cells using the pET-24a vector, two recombinant constructs generated by site-directed mutagenesis of the IPNV VP4 protease analyzed
wild-type IPNA and truncated forms, expressed in Escherichia coli
expression in Escherichia coli JM107
-
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Petit, S.; Lejal, N.; Huet, J.C.; Delmas, B.
Active residues and viral substrate cleavage sites of the protease of the birnavirus infectious pancreatic necrosis virus
J. Virol.
74
2057-2066
2000
infectious pancreatic necrosis virus (Q703G9)
Manually annotated by BRENDA team
Gorbalenya, A.E.; Mundt, E.
Birnavirus VP4 processing endopeptidase
Handbook of Proteolytic Enzymes (Barrett, A. J. , Rawlings, N. D. , Woessner, J. F. , eds. )Academic Press
2
1995-1998
2004
Blotched snakehead virus, Drosophila X virus, Infectious bursal disease virus, infectious pancreatic necrosis virus
-
Manually annotated by BRENDA team
Manning, D.S.; Leong, J.C.
Expression in Escherichia coli of the large genomic segment of infectious pancreatic necrosis virus
Virology
179
16-25
1990
infectious pancreatic necrosis virus
Manually annotated by BRENDA team
Lee, J.; Feldman, A.R.; Delmas, B.; Paetzel, M.
Crystal structure of the VP4 protease from infectious pancreatic necrosis virus reveals the acyl-enzyme complex for an intermolecular self-cleavage reaction
J. Biol. Chem.
282
24928-24937
2007
infectious pancreatic necrosis virus (Q703G9)
Manually annotated by BRENDA team