Information on EC 3.4.24.67 - choriolysin H

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
3.4.24.67
-
RECOMMENDED NAME
GeneOntology No.
choriolysin H
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
Hydrolysis of the inner layer of fish egg envelope. Also hydrolysis of casein and small molecule substrates such as succinyl-Leu-Leu-Val-Tyr-/-7-(4-methyl)coumarylamide
show the reaction diagram
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-
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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
177529-16-7
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
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-
-
Manually annotated by BRENDA team
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SwissProt
Manually annotated by BRENDA team
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-
-
Manually annotated by BRENDA team
flounder
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-
Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
Benzoyl-Arg 4-methylcoumarin 7-amide + H2O
?
show the reaction diagram
-
hydrolysis at about 3% the rate of succinyl-Leu-Leu-Val-Tyr 4-methylcoumarin 7-amide
-
-
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benzyloxycarbonyl-Ala-Ala-Asn-4-methylcoumarin 7-amide + H2O
?
show the reaction diagram
-
about 8.5% of the rate with benzyloxycarbonyl-Phe-Arg-4-methylcoumarin 7-amide
-
-
?
benzyloxycarbonyl-Phe-Arg-4-methylcoumarin 7-amide + H2O
benzyloxycarbonyl-Phe + Arg-4-methylcoumarin 7-amide
show the reaction diagram
-
-
-
-
?
Benzyloxycarbonyl-Phe-Ser-Arg 4-methylcoumarin 7-amide + H2O
?
show the reaction diagram
-
hydrolysis at about 2% the rate of succinyl-Leu-Leu-Val-Tyr 4-methylcoumarin 7-amide
-
-
-
benzyloxycarbonyl-Val-Pro-Arg-4-methylcoumaryl-7-amide + H2O
?
show the reaction diagram
-
preferred by HCE1
-
-
?
casein + H2O
?
show the reaction diagram
chorion + H2O
?
show the reaction diagram
Chorion + H2O
Swollen chorion + ?
show the reaction diagram
EVLPLDNPPPA + H2O
EVLPLD + NPPPA
show the reaction diagram
GSKTPQDPTQQ + H2O
GSKTPQ + DPTQQ
show the reaction diagram
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location: Q39/D40 in ChgL (ZI-3). Specific activity: 28.5 nmol/30min/microgram enzyme
-
-
?
KLMLKAPEPF + H2O
KLMLK + APEPF
show the reaction diagram
-
synthetic peptide substrate designed according to cleavage site in egg envelope protein. Cleavage at 7.6% of the rate with TVQQSDYLIK
-
-
?
N-tert-Butoxycarbonyl-Val-Pro-Arg 4-methylcoumarin 7-amide + H2O
?
show the reaction diagram
-
hydrolysis at about 12% the rate of succinyl-Leu-Leu-Val-Tyr 4-methylcoumarin 7-amide
-
-
-
NPQVPQYPSKPQ + H2O
NPQVPQ + YPSKPQ
show the reaction diagram
NPSYPQNPSYPQ + H2O
NPSYPQ + NPSYPQ
show the reaction diagram
Peptidyl 4-methylcoumarin 7-amides + H2O
?
show the reaction diagram
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i.e. MCA-peptides
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-
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PGKNPNTPPIG + H2O
PGKNPN + TPPIG
show the reaction diagram
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location: N220/T221 in ChgH (ZI-1,2). Specific activity: 11.3 nmol/30min/microgram enzyme
-
-
?
PLPVRVEEVV + H2O
PLPVR + VEEVV
show the reaction diagram
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synthetic peptide substrate designed according to cleavage site in egg envelope protein. Cleavage at 38% of the rate with TVQQSDYLIK
-
-
?
PSKRPEAPGVP + H2O
PSKRPE + APGVP
show the reaction diagram
QTPSYPQQ + H2O
QTPS + YPQQ
show the reaction diagram
SAPVVRTSQAV + H2O
NPQVPQ + YPSKPQ
show the reaction diagram
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synthetic substrate generated from the 25 kDa product in hatching liquid with an inefficient cleavage site
-
-
?
SKPQYPNP + H2O
SKPQ + YPNP
show the reaction diagram
Suc-Leu-Leu-Val-Tyr-4-methylcoumarin-7-amide + H2O
?
show the reaction diagram
-
-
-
-
?
Succinyl-Ala-Ala-Pro-Phe 4-methylcoumarin 7-amide + H2O
?
show the reaction diagram
-
hydrolysis at about 13% the rate of succinyl-Leu-Leu-Val-Tyr 4-methylcoumarin 7-amide
-
-
-
Succinyl-Ala-Pro-Ala 4-methylcoumarin 7-amide + H2O
?
show the reaction diagram
-
hydrolysis at about 45% the rate of succinyl-Leu-Leu-Val-Tyr 4-methylcoumarin 7-amide
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-
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succinyl-Ala-Pro-Ala-4-methylcoumaryl-7-amide + H2O
?
show the reaction diagram
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best substrate for both isoforms of HCE
-
-
?
succinyl-Ile-Ile-Trp-4-methylcoumarin 7-amide + H2O
?
show the reaction diagram
-
about 5.7% of the rate with benzyloxycarbonyl-Phe-Arg-4-methylcoumarin 7-amide
-
-
?
Succinyl-Leu-Leu-Val-Tyr 4-methylcoumarin 7-amide + H2O
Succinyl-Leu-Leu-Val-Tyr + 7-amino-4-methylcoumarin
show the reaction diagram
-
best substrate of peptidyl 4-methylcoumarin 7-amides
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-
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succinyl-Leu-Leu-Val-Tyr-4-methylcoumarin 7-amide + H2O
?
show the reaction diagram
-
about 40% of the rate with benzyloxycarbonyl-Phe-Arg-4-methylcoumarin 7-amide
-
-
?
succinyl-Leu-Leu-Val-Tyr-4-methylcoumaryl-7-amide + H2O
?
show the reaction diagram
-
preferred by HCE2
-
-
?
t-butyloxycarbonyl-Phe-Ser-Arg-4-methylcoumarin 7-amide + H2O
?
show the reaction diagram
-
about 9% of the rate with benzyloxycarbonyl-Phe-Arg-4-methylcoumarin 7-amide
-
-
?
TVQQSDYLIK + H2O
TVQQS + DYLIK
show the reaction diagram
-
synthetic peptide substrate designed according to cleavage site in egg envelope protein
-
-
?
VPFELRYPVPA + H2O
VPFELR + YPVPA
show the reaction diagram
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specific activity: 0.05 nmol/min/microgram protein
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-
?
YPSKPQTPTET + H2O
YPSKPQ + TPTET
show the reaction diagram
ZP2 + H2O
?
show the reaction diagram
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zebrafish egg envelope glycoprotein, cleavage at Ser47/Asp48 and Arg66/Val67 in the unfertilized egg, at Gln79/Ala80 in the fertilized egg
-
-
?
ZP3 + H2O
?
show the reaction diagram
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zebrafish egg envelope glycoprotein, cleavage at Gln79/Ala80 in the unfertilized egg, at Gln67/Ala68 in the fertilized egg
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-
?
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
chorion + H2O
?
show the reaction diagram
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Calcium
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0.00286-0.00296 mg/mg protein, inductively coupled plasma emission analysis
Cu2+
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high concentrations reactivate EDTA-inactivated apoenzyme
Magnesium
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0.00063-0.00076 mg/mg protein, inductively coupled plasma emission analysis
Zinc
-
0.00127-0.00143 mg/mg protein, inductively coupled plasma emission analysis
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Anti-HCE antibody
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inhibits choriolytic swelling and binding to chorion, no effect on caseinolytic activity
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antichoriolysin H
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inhibits both choriolytic activity and binding to the egg envelope, does not affect caseinolytic activity
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chymostatin
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Cu2+
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about 95% inhibition at 5 mM
iodoacetamide
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lima bean trypsin-chymotrypsin inhibitor
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LBTI
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ovomucoid
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pepstatin A
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soybean trypsin-chymotrypsin inhibitor
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SBTI
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additional information
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
4.28
casein
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pH 7.0, 35°C
480
NPQVPQYPSKPQ
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pH 8.0, 30°C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
8 - 8.7
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for choriolytic and caseinolytic activity, respectively
8.7
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choriolytic activity
additional information
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pI: 10.2
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.5 - 8.9
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about 50% of maximal caseinolytic activity at pH 6.5 and 8.9
7.9 - 9
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about 50% of maximal choriolytic activity at pH 7.9 and 9
additional information
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pH profile
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
-
temperature profile
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
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in stage 25 embryos, strong expression of HCE found in branchial arches
Manually annotated by BRENDA team
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in stage 21 embryos, the HCE-expressing cells were located between two eye rudiments
Manually annotated by BRENDA team
ovarian fluid carrying the embryos immediately before the hatching stage; ovarian fluid carrying the embryos immediately before the hatching stage
Manually annotated by BRENDA team
additional information
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first expressed in the pillow anterior to the forebrain of early neurula
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
immunocytochemical detection
Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
20000
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Oryzias latipes, deduced from sedimentation coefficient
22584
x * 22584, calculated, x * 23000, SDS-PAGE, x * 22789, MALDI-TOF/MS
22789
x * 22584, calculated, x * 23000, SDS-PAGE, x * 22789, MALDI-TOF/MS
22900
Oryzias latipes, deduced from nucleotide sequence of clone HCE21
23000
x * 22584, calculated, x * 23000, SDS-PAGE, x * 22789, MALDI-TOF/MS; x * 23 056, calculated, x * 23000, SDS-PAGE, x * 23075, MALDI-TOF/MS
23010
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Oryzias latipes, HCE-1, estimation on the basis of amino acid composition
23075
x * 23 056, calculated, x * 23000, SDS-PAGE, x * 23075, MALDI-TOF/MS
23470
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Oryzias latipes, HCE-2, estimation on the basis of amino acid composition
34800
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x * 34800, SDS-PAGE
additional information
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amino acid composition of isomers HCE-1 and HCE-2
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
hanging drop vapour-diffusion method, crystals diffracted to 1.34 A resolution
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TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
10 - 40
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10 min, gradual decrease of choriolytic activity (10-30% loss)
25
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4 days, in 50 mM Tris-HCl, 10 mM NaCl, pH 7.5, about 80% loss of choriolytic activity, 0.05% Brij-35 stabilizes, caseinolytic activity is less susceptible to long-term incubation
44
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10 min, about 50% loss of choriolytic activity
50
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10 min, about 90% loss of choriolytic activity
60
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inactivation
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
0°C, in the presence of Ca2+ and Mg2+ in addition to Zn2+, 9 weeks
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
2 isomers: HCE-1 and HCE-2, separable by HPLC ion exchange chromatography; from hatching liquid
-
cationic column chromatography
-
from hatching liquid
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gel filtration, cation exchange column chromatography
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native enzyme 40fold from embryos by gel filtration and anion exchange chromatography
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partly purified by cation-exchange column chromatography and gel filtration, two isoforms
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
five copies of MHCE gene including one pseudogene, DNA and amino acid sequence determination and analysis, genetic structure, genes for hatching enzyme MHCE are intron-less, sequence comparison and phylogenetic analysis
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into the pGEM-T vector system
O2 not identical but closely similar clones HCE21 and HCE23
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
degradation
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HCE dissolves the inner layer of the egg envelope to facilitate hatching of the embryo