Information on EC 3.4.22.B68 - EhCP4 proteinase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
3.4.22.B68
preliminary BRENDA-supplied EC number
RECOMMENDED NAME
GeneOntology No.
EhCP4 proteinase
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
unique preference for valine and isoleucine at P2 and broader specificities at the P1 and P3 positions, while proline at P4 is relatively selective
show the reaction diagram
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-
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
strain HM1-IMSS
UniProt
Manually annotated by BRENDA team
strain HM1-IMSS
UniProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
physiological function
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CP4 is a virulence factor involved in parasite adherence, cell-detaching, hemolysis and inducing apoptosis of human vaginal epithelial cells
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
benzyloxycarbonyl-VVR-7-amido-4-methylcoumarin + H2O
benzyloxycarbonyl-VVR + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
?
complement component C3 + H2O
?
show the reaction diagram
immunglobulin A + H2O
?
show the reaction diagram
-
-
-
?
immunglobulin G + H2O
?
show the reaction diagram
-
-
-
?
Laminin-1 + H2O
?
show the reaction diagram
pro-interleukin-18 + H2O
?
show the reaction diagram
-
-
-
?
Protein + H2O
?
show the reaction diagram
succinyl-LLVY-7-amido-4-methylcoumarin + H2O
succinyl-LLVY + 7-amino-4-methylcoumarin
show the reaction diagram
villin-1 + H2O
?
show the reaction diagram
-
-
-
?
additional information
?
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NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
complement component C3 + H2O
?
show the reaction diagram
immunglobulin A + H2O
?
show the reaction diagram
C4LTT0
-
-
-
?
immunglobulin G + H2O
?
show the reaction diagram
C4LTT0
-
-
-
?
Laminin-1 + H2O
?
show the reaction diagram
pro-interleukin-18 + H2O
?
show the reaction diagram
C4LTT0
-
-
-
?
villin-1 + H2O
?
show the reaction diagram
C4LTT0
-
-
-
?
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
WRR605
CP4-specific inhibitor, synthesized based on the substrate specificity of CP4, inhibits the recombinant enzyme in vitro
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.03 - 0.078
benzyloxycarbonyl-VVR-7-amido-4-methylcoumarin
0.031
succinyl-LLVY-7-amido-4-methylcoumarin
recombinant enzyme, at pH 7.0, 37°C
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00967
WRR605
apparent value, pH 7.0, at 37°C
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7
the optimum for substrate cleavage is at pH 7.0
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
23000
calculated molecular mass of the mature enzyme
26000
active recombinant CP4, SDS-PAGE
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
3 - 4
recombinant CP4 is efficiently autoactivated at pH 3.0-4.0 with 5 mM dithiothreitol, mature recombinant CP4 is unstable at acid pH and undergoes further degradation
709110
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
nickel affinity column chromatography
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli BL21 Condon Plus (DE3)-RIPL cells as as a fusion protein with bacterial thioredoxin A with the His tag at the amino terminus of the prodomain
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
co-culture of Entamoeba histolytica trophozoites with mucin-producing T84 cells increases ehcp4 expression up to 6fold
Renatured/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
maximal active enzyme is obtained by reducing the denatured thioredoxin A-CP4 fusion protein with 25 mM DTT at 37°C for 1 h followed by rapid dilution of the protein into ice-cold refolding buffer (100 mM Tris-HCl pH 8.5, 100 mM NaCl, 20% (v/v) glycerol, 250 mM arginine-HCl, 2 mM EDTA, pH 8, 10 mM GSH, 1 mM L-GSSG disodium salt)