Information on EC 3.4.24.55 - pitrilysin

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The expected taxonomic range for this enzyme is: Bacteria

EC NUMBER
COMMENTARY hide
3.4.24.55
-
RECOMMENDED NAME
GeneOntology No.
pitrilysin
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
Preferential cleavage of -Tyr16-/- Leu- and -Phe25-/- Tyr-bonds of oxidized insulin B chain. Also acts on other substrates of less than 7 kDa such as insulin and glucagon
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of peptide bond
-
-
-
-
CAS REGISTRY NUMBER
COMMENTARY hide
81611-78-1
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
strain H4
-
-
Manually annotated by BRENDA team
strain H4
-
-
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
(7-Methoxycoumarin-4-yl)acetyl-Nle-Ala-Val-Lys-Tyr-Leu-Asn-Ser-(2,4-dinitrophenyl)Lys-Leu-Asp-D-Lys + H2O
?
show the reaction diagram
-
-
-
-
-
amyloid beta-peptide + H2O
?
show the reaction diagram
-
-
-
-
?
amyloid beta-peptide with mutations K16A/K28A + H2O
?
show the reaction diagram
-
cleavage site is identical to that of native amyloid beta-peptide
-
-
?
Angiotensinogen(1-14) + H2O
?
show the reaction diagram
-
-
-
-
-
beta-amyloid + H2O
?
show the reaction diagram
-
18% of the activity with beta-endorphin, major cleavage site His14-Gln15
-
-
?
beta-endorphin + H2O
?
show the reaction diagram
-
cleavage at Lys19-Asn20 bond. Residues Leu14, Val15 and Leu17 and region 22-26 are responsible for recognition by enzyme
-
-
?
calcitonin + H2O
?
show the reaction diagram
-
3% of the activity with beta-endorphin
-
-
?
dynorphin + H2O
?
show the reaction diagram
dynorphin A fragment 1-13 + H2O
?
show the reaction diagram
Glucagon + H2O
?
show the reaction diagram
Insulin + H2O
Hydrolyzed insulin
show the reaction diagram
Insulin B-chain + H2O
?
show the reaction diagram
Insulin-like growth factor II + H2O
?
show the reaction diagram
-
-
-
-
-
Secretin + H2O
?
show the reaction diagram
-
-
-
-
-
Substance P + H2O
?
show the reaction diagram
-
10% of the activity with beta-endorphin
-
-
?
Thyrocalcitonin + H2O
?
show the reaction diagram
-
-
-
-
-
Vasoactive intestinal peptide + H2O
?
show the reaction diagram
-
7.2% of the activity with beta-endorphin, major cleavage site Arg14-Lys15
-
-
?
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ca2+
-
reactivation after inhibition with chelating agents
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1,10-phenanthroline
2,6-pyridinedicarboxylic acid
-
-
2-mercaptoethanol
-
1% w/v, 95% inhibition
bacitracin
Ca2+
-
1 mM, weak, reactivates after inhibition with chelating agents
Chelating agents
-
-
Cu2+
-
1 mM, complete inhibition
cysteine
-
1 mM, complete inhibition
dynorphin A(1-13)
-
-
E-64
-
1 mM, 46% inhibition
EGTA
-
1 mM, complete inhibition
Fe2+
-
1 mM, 76% inhibition
Mn2+
-
1 mM, weak, reactivates after inhibition with chelating agents
N-ethylmaleimide
-
1 mM, 95% inhibition
phosphoramidon
-
-
Tetraethylenepentamine
-
-
Zincov
-
-
additional information
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0077
(7-methoxycoumarin-4-yl)acetyl-Nle-Ala-Val-Lys-Tyr-Leu-Asn-Ser-Lys(2,4-dinitrophenyl)-Leu-Asp-D-Lys
-
-
0.0009
beta-amyloid
-
pH 8.0, 37C
-
0.00036
beta-endorphin
-
pH 8.0, 37C
0.0022
calcitonin
-
pH 8.0, 37C
0.12
dynorphin A fragment 1-13
-
pH 8.0, 37C
0.00061 - 0.0127
glucagon
0.000105
Insulin
-
-
-
0.0015 - 0.05
Insulin B-chain
0.031
Substance P
-
pH 8.0, 37C
0.0004
Vasoactive intestinal peptide
-
pH 8.0, 37C
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2.9
(7-Methoxycoumarin-4-yl)acetyl-Nle-Ala-Val-Lys-Tyr-Leu-Asn-Ser-(2,4-dinitrophenyl)Lys-Leu-Asp-D-Lys
Escherichia coli
-
-
2.33
beta-amyloid
Escherichia coli
-
pH 8.0, 37C
-
12.5
beta-endorphin
Escherichia coli
-
pH 8.0, 37C
0.43
calcitonin
Escherichia coli
-
pH 8.0, 37C
0.55
dynorphin A fragment 1-13
Bacillus halodurans
-
pH 8.0, 37C
0.05
glucagon
Escherichia coli
-
-
0.18 - 2.66
Insulin B-chain
0.63
Substance P
Escherichia coli
-
pH 8.0, 37C
0.9
Vasoactive intestinal peptide
Escherichia coli
-
pH 8.0, 37C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
-
-
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.8 - 7.2
-
insulin
7.5
-
4-dinitrophenyl)Lys-Leu-Asp-D-Lys
8
-
activity in sodium phosphate buffer is 3- to 4fold higher than in veronal or Tris/HCl buffer
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.4
-
calculated
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
PDB
SCOP
CATH
ORGANISM
UNIPROT
Escherichia coli (strain K12)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
46500
-
gel filtration
107700
-
E. coli, calculation from amino acid sequence
108000
-
Acinetobacter calcoaceticus, gel filtration
112000
-
Acinetobacter calcoaceticus, native electrophoresis
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
tetramer
-
4 * 27000, Acinetobacter calcoaceticus, SDS-PAGE
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
40
-
10 min, stable
50
-
half-life is 20 min
55
-
half-life is 10 min
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20C, 50 mM Tris-HCl, pH 7.5, 40% v/v glycerol
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
recombinant protein with His-tag
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D79A
-
no effect on function
E91A
-
no proteolytic activity
F198A
-
enzyme remains functional
H193A
-
enzyme remains functional
K197A
-
enzyme remains functional
N119A
-
promotion of yeast mating factor in heterologous expression is disabled
N317A/R318A/S319A
-
enzyme remains functional
R792A
-
no proteolytic activity
R792A/T793A/E794A/E795A
-
no proteolytic activity
S117A
-
enzyme remains functional
S117A/H118A/N119A
-
promotion of yeast mating factor in heterologous expression is disabled
S196A/K197A/F198A/S199A
-
promotion of yeast mating factor in heterologous expression is disabled
T127A
-
enzyme remains functional
T322A/L323A
-
enzyme remains functional
Y799A
-
no proteolytic activity
additional information
-
heterologous expression of enzyme in yeast, enzyme functionally substitutes for yeast Ax11p and Ste23p in pheromone production. Independent heterologous expression of N-terminal domain with amino acids 1-515 or C-terminal domain with amino acids 516-962 does not promote yeast mating. Co-expression of both domains restores yeast mating
Show AA Sequence (726 entries)
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