Information on EC 3.4.11.4 - tripeptide aminopeptidase

Word Map on EC 3.4.11.4
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Select one or more organisms in this record:
Show additional data
Do not include text mining results
Include (text mining) results (more...)
Include results (AMENDA + additional results, but less precise; more...)


The expected taxonomic range for this enzyme is: Eukaryota, Bacteria

EC NUMBER
COMMENTARY hide
3.4.11.4
-
RECOMMENDED NAME
GeneOntology No.
tripeptide aminopeptidase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
release of the N-terminal residue from a tripeptide
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis
-
-
hydrolysis of peptide bond
-
-
-
-
CAS REGISTRY NUMBER
COMMENTARY hide
9056-26-2
-
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(Gln)17-Arg-Arg-Gly-Arg-Arg + H2O
(Gln)14-Arg-Arg-Gly-Arg-Arg + Gln-Gln-Gln
show the reaction diagram
-
little activity
-
-
?
(Gln)20-Arg-Arg-Gly-Arg-Arg + H2O
(Gln)17-Arg-Arg-Gly-Arg-Arg + Gln-Gln-Gln
show the reaction diagram
-
little activity
-
-
?
Ala-Ala-Ala + H2O
Ala + Ala-Ala
show the reaction diagram
26% of the activity with Met-Gly-Gly
-
?
Ala-Ala-Phe-4-methylcoumarin 7-amide + H2O
?
show the reaction diagram
-
-
-
-
?
Ala-Ala-Phe-7-amido-4-methylcoumarin + H2O
?
show the reaction diagram
Ala-Ala-Phe-7-amido-4-methylcoumarin + H2O
Ala + Ala-Phe-7-amido-4-methylcoumarin
show the reaction diagram
Ala-Ala-Phe-7-amido-4-methylcoumarin + H2O
Ala-Ala-Phe + 7-amino-4-methylcoumarin
show the reaction diagram
Ala-Ala-Phe-p-nitroanilide + H2O
Ala-Ala-Phe + p-nitroaniline
show the reaction diagram
-
-
-
?
Ala-Phe-Pro-beta-naphthylamide + H2O
Ala-Phe-Pro + beta-naphthylamine
show the reaction diagram
Angiotensin II + H2O
?
show the reaction diagram
-
-
-
-
?
angiotensin III + H2O
?
show the reaction diagram
-
-
-
-
?
Arg-Ala-Arg + H2O
Arg + Ala-Arg
show the reaction diagram
-
-
-
-
?
Arg-Ser-Arg + H2O
Arg + Ser-Arg
show the reaction diagram
-
-
-
-
?
casein + H2O
?
show the reaction diagram
-
assay at pH 8.0, 37C
-
-
?
cholecystokinin octapeptide + H2O
?
show the reaction diagram
-
-
-
-
?
des-Tyr-dynorphin A1-8 + H2O
?
show the reaction diagram
-
-
-
-
?
enkephalin + H2O
?
show the reaction diagram
-
-
-
-
?
Gly-7-amido-4-methylcoumarin + H2O
7-amino-4-methylcoumarin + Gly
show the reaction diagram
-
assay at pH 8.0, 37C
-
-
?
Gly-Ala-Pro-beta-naphthylamide + H2O
Gly-Ala-Pro + beta-naphthylamine
show the reaction diagram
Gly-Gly-Phe + H2O
Gly + Gly-Phe
show the reaction diagram
Gly-Leu-Tyr + H2O
Gly + Leu-Tyr
show the reaction diagram
L-Ala-L-Ala-D-Ala + H2O
L-Ala + L-Ala-D-Ala
show the reaction diagram
-
-
-
-
?
L-Ala-L-Ala-L-Ala + H2O
L-Ala + L-Ala-L-Ala
show the reaction diagram
L-Ala-L-Ala-L-Phe-7-amido-4-methylcoumarin + H2O
7-amino-4-methylcoumarin + L-Ala-L-Ala-L-Phe
show the reaction diagram
-
assay at pH 8.0, 37C
-
-
?
L-Asp-7-amido-4-methylcoumarin + H2O
7-amino-4-methylcoumarin + L-Asp
show the reaction diagram
-
assay at pH 8.0, 37C
-
-
?
L-Gln-7-amido-4-methylcoumarin + H2O
7-amino-4-methylcoumarin + L-Gln
show the reaction diagram
-
assay at pH 8.0, 37C
-
-
?
L-Leu-7-amido-4-methylcoumarin + H2O
7-amino-4-methylcoumarin + L-Leu
show the reaction diagram
-
assay at pH 8.0, 37C
-
-
?
L-Lys-4-nitroanilide + H2O
4-nitroaniline + L-Lys
show the reaction diagram
-
assay at pH 8.0, 37C
-
-
?
L-Met-7-amido-4-methylcoumarin + H2O
7-amino-4-methylcoumarin + L-Met
show the reaction diagram
-
assay at pH 8.0, 37C
-
-
?
L-Met-L-Met-L-Met + H2O
L-Met + L-Met-L-Met
show the reaction diagram
L-Phe-Gly-Gly + H2O
L-Phe + Gly-Gly
show the reaction diagram
L-Pro-7-amido-4-methylcoumarin + H2O
7-amino-4-methylcoumarin + L-Pro
show the reaction diagram
-
assay at pH 8.0, 37C
-
-
?
L-Thr-7-amido-4-methylcoumarin + H2O
7-amino-4-methylcoumarin + L-Thr
show the reaction diagram
-
assay at pH 8.0, 37C
-
-
?
L-Trp-7-amido-4-methylcoumarin + H2O
7-amino-4-methylcoumarin + L-Trp
show the reaction diagram
-
assay at pH 8.0, 37C
-
-
?
L-Tyr-7-amido-4-methylcoumarin + H2O
7-amino-4-methylcoumarin + L-Tyr
show the reaction diagram
-
assay at pH 8.0, 37C
-
-
?
Leu-Gly-Gly + H2O
Leu + Gly-Gly
show the reaction diagram
Leu-Leu-Ala + H2O
Leu + Leu-Ala
show the reaction diagram
33% of the activity with Met-Gly-Gly
-
?
Leu-Leu-Leu + H2O
Leu + Leu-Leu
show the reaction diagram
15% of the activity with Met-Gly-Gly
-
?
Lys-Gly-Gly + H2O
Lys + Gly-Gly
show the reaction diagram
13% of the activity with Met-Gly-Gly
-
?
Met-Ala-Ser + H2O
?
show the reaction diagram
-
-
-
-
?
Met-Ala-Ser + H2O
Met + Ala-Ser
show the reaction diagram
-
-
-
?
Met-Gly-Gly + H2O
Met + Gly-Gly
show the reaction diagram
microcin C + H2O
?
show the reaction diagram
-
microcin C is processed to a nonhydrolyzable analog of aspartyl-adenylate
-
-
?
neurokinin alpha + H2O
?
show the reaction diagram
-
-
-
-
?
oxidized insulin B chain + H2O
?
show the reaction diagram
-
assay at pH 8.0, 37C
-
-
?
Ser-Ser-Ser + H2O
?
show the reaction diagram
-
-
-
-
?
tripeptides + H2O
?
show the reaction diagram
-
e.g. Met-Leu-Tyr: chemotactic factor, Gly-His-Lys: liver growth factor, slow hydrolysis
-
-
-
Tyr-Gly-Gly + H2O
Tyr + Gly-Gly
show the reaction diagram
11% of the activity with Met-Gly-Gly
-
?
additional information
?
-
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
tripeptides + H2O
?
show the reaction diagram
-
e.g. Met-Leu-Tyr: chemotactic factor, Gly-His-Lys: liver growth factor, slow hydrolysis
-
-
-
additional information
?
-
-
the enzyme digests peptide products of the 26S proteasome and other endopeptidases into tripeptides
-
-
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ca2+
-
stimulates
additional information
-
no metalloenzyme
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1,10-phenanthroline
2,5-diamino-8-carbamimidamido-N-(4,8-diamino-5-methyl-1-methylidene-2,3,7-trioxononyl)-4-oxooctanamide
-
potent inhibitor
2-mercaptoethanol
5,5'-dithiobis(2-nitrobenzoate)
-
-
5-5'-dithiobis(2-nitrobenzoic acid)
-
-
Ala-Ala-Phe chloromethyl ketone
-
-
Ala-Ala-Phe-chloromethyl ketone
-
85-90% inhibition at 1 mM
Ala-Ala-Phe-chloromethylketone
Ala-Ile-pyrrolidin-2-yl boronic acid
-
-
Amino aldehydes
-
e.g. leucinal, prolinal, alaninal
antipain
-
-
bacitracin
-
-
bestatin
butabindide
Carboxy-o-phenanthroline
-
-
chymostatin
-
-
D-3-thio-2-methylpropanyl proline
-
-
diisopropylfluorophosphate
-
strongly inhibitory at 1 mM
dithiothreitol
iodoacetate
-
-
iodoacetic acid
-
partial inhibition at 1 mM
L-Leu-Gly-Gly
-
substrate inhibition
Met-enkephalin
-
-
Met-Leu-Tyr
-
-
molybdate
-
-
N-ethylmaleimide
o-phenanthroline
-
-
O3-tertiary-butyl-threonylphenylalanylproline
-
-
-
p-chloromercuribenzoate
-
-
p-Chloromercuriphenyl sulfonic acid
-
-
p-Chloromercuriphenylsulfonic acid
-
strongly inhibitory at 1 mM
p-hydroxymercuribenzoate
p-Hydroxymercuriphenyl sulfonate
-
-
Pepstatin
Phenylmethanesulfonylfluoride
-
-
phenylmethylsulfonyl fluoride
0.1 mM, 89% inhibition
puromycin
-
-
tosyl-L-lysine-chloromethylketone
-
-
tosyl-phenylalanine-chloromethylketone
-
-
Tripeptides with Pro in N-terminal position
-
-
Z-Gly-Leu-Ala-OH
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2-mercaptoethanol
-
activates
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.52
Ala-Ala-Ala
-
-
1.4
Ala-Ala-D-Ala
-
-
0.076 - 0.25
Ala-Ala-Phe-7-amido-4-methylcoumarin
1.4
Ala-Gly-Gly
-
-
0.17 - 0.85
Ala-Phe-Pro-beta-naphthylamide
0.0065
Arg-Ala-Arg
-
calculated value, wild type enzyme
0.0012
Arg-Ser-Arg
-
calculated value, wild type enzyme
0.16
Gly-7-amido-4-methylcoumarin
-
pH 8.0, 37C
0.38 - 0.42
Gly-Ala-Pro-beta-naphthylamide
7.5 - 20
Gly-Gly-Gly
4.91 - 13
Gly-Gly-Phe
0.33
L-Ala-L-Ala-L-Phe-7-amido-4-methylcoumarin
-
pH 8.0, 37C
0.401
L-Asp-7-amido-4-methylcoumarin
-
pH 8.0, 37C
0.14
L-Leu-7-amido-4-methylcoumarin
-
pH 8.0, 37C
0.17
L-Lys-4-nitroanilide
-
pH 8.0, 37C
0.14
L-Met-7-amido-4-methylcoumarin
-
pH 8.0, 37C
0.25 - 0.6
Leu-Gly-Gly
3.3 - 5.1
Met-Ala-Ser
2.6 - 5.3
Met-Gly-Gly
0.33 - 1.1
Phe-Gly-Gly
1.3
Pro-Gly-Gly
-
-
additional information
additional information
-
-
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.04 - 41.66
Ala-Ala-Phe-7-amido-4-methylcoumarin
6.8 - 511
Ala-Phe-Pro-beta-naphthylamide
5.3
Arg-Ala-Arg
Homo sapiens
-
calculated value, wild type enzyme
0.56
Arg-Ser-Arg
Homo sapiens
-
calculated value, wild type enzyme
12
Gly-7-amido-4-methylcoumarin
Escherichia coli
-
pH 8.0, 37C
265 - 354
Gly-Ala-Pro-beta-naphthylamide
2030
Gly-Gly-Gly
Sus scrofa
-
-
14.4
L-Ala-L-Ala-L-Phe-7-amido-4-methylcoumarin
Escherichia coli
-
pH 8.0, 37C
11.7
L-Asp-7-amido-4-methylcoumarin
Escherichia coli
-
pH 8.0, 37C
19
L-Leu-7-amido-4-methylcoumarin
Escherichia coli
-
pH 8.0, 37C
14.3
L-Lys-4-nitroanilide
Escherichia coli
-
pH 8.0, 37C
14
L-Met-7-amido-4-methylcoumarin
Escherichia coli
-
pH 8.0, 37C
additional information
additional information
Bacillus subtilis
-
-
-
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
71
Gly-7-amido-4-methylcoumarin
Escherichia coli
-
pH 8.0, 37C
8818
41
L-Ala-L-Ala-L-Phe-7-amido-4-methylcoumarin
Escherichia coli
-
pH 8.0, 37C
10236
23
L-Asp-7-amido-4-methylcoumarin
Escherichia coli
-
pH 8.0, 37C
25184
112
L-Leu-7-amido-4-methylcoumarin
Escherichia coli
-
pH 8.0, 37C
3540
89
L-Lys-4-nitroanilide
Escherichia coli
-
pH 8.0, 37C
3734
104
L-Met-7-amido-4-methylcoumarin
Escherichia coli
-
pH 8.0, 37C
3910
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00002
2,5-diamino-8-carbamimidamido-N-(4,8-diamino-5-methyl-1-methylidene-2,3,7-trioxononyl)-4-oxooctanamide
-
-
0.0000881 - 0.0488
Ala-Ile-pyrrolidin-2-yl boronic acid
0.00021
butabindide
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
1.7
-
value about, substrate L-Ala-L-Ala-L-Phe-7-amido-4-methylcoumarin; value about, substrate L-Gln-7-amido-4-methylcoumarin; value about, substrate L-Thr-7-amido-4-methylcoumarin; value about, substrate L-Trp-7-amido-4-methylcoumarin
2.5
-
value about, substrate L-Tyr-7-amido-4-methylcoumarin
3.3
-
value about, substrate L-Pro-7-amido-4-methylcoumarin
8.3
-
value about, substrate Gly-7-amido-4-methylcoumarin
16.7
-
value about, substrate L-Asp-7-amido-4-methylcoumarin
25
-
value about, substrate L-Met-7-amido-4-methylcoumarin
31.7
-
value about, substrate L-Lys-4-nitroanilide
35
-
value about, substrate L-Leu-7-amido-4-methylcoumarin
59.5
-
-
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.4
-
and pH 7.6
7.7
-
in presence of 2 mM dithiothreitol
7.8 - 8
-
-
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.5 - 7.2
-
more than 50% activity
6.8 - 10
-
more than 50% activity
9
-
no activity above
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30
-
assay at
37
-
assay at
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
tritosomes
Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
45000
-
2 * 45000, SDS-PAGE
47000
3 * 47000, SDS-PAGE
47500
-
2 * 47500, SDS-PAGE
50000 - 57000
-
ultracentrifugation, SDS-PAGE
51400
-
gel filtration
55000
-
gel filtration
57000
-
1 * 57000, SDS-PAGE
58000
-
4 * 58000, SDS-PAGE
65000
-
gel filtration
68000
-
gel filtration
71000
-
peptidase-1, sedimentation equilibrium centrifugation
77000
-
2 * 77000, SDS-PAGE
79390
-
calculated from amino acid sequence
80000
-
gel filtration
80900
-
2 * 80900, tripeptidase-2, sedimentation equilibrium centrifugation in 6 M guanidium chloride + mercaptoethanol
100000
108000
-
gel filtration
137200
-
tripeptidase-2, sedimentation equilibrium centrifugation
138000
-
x * 138000
140000
-
40 * 140000
142000
-
x * 142000, the 142000 Da enzyme form differs from the 153000 Da enzyme mainly by a truncation at the C-terminal end, SDS-PAGE
145000
-
gel filtration
150000
gel filtration
153000
-
x * 153000, the 142000 Da enzyme form differs from the 153000 Da enzyme mainly by a truncation at the C-terminal end, SDS-PAGE
154000
-
SDS-PAGE
230000
-
gel filtration
275000
-
SDS-PAGE
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
-
x * 142000, the 142000 Da enzyme form differs from the 153000 Da enzyme mainly by a truncation at the C-terminal end, SDS-PAGE; x * 153000, the 142000 Da enzyme form differs from the 153000 Da enzyme mainly by a truncation at the C-terminal end, SDS-PAGE
hexamer
-
-
homodimer
-
2 * 77000, SDS-PAGE
homooligomer
monomer
tetramer
-
4 * 58000, SDS-PAGE
trimer
3 * 47000, SDS-PAGE
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
no modification
-
no glycoprotein
side-chain modification
-
glycoprotein
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
hanging drop vapour diffusion method with 100 mM Ches (pH 9.0), 1.1 M potassium/sodium tartrate, 200 mM lithium sulfate
-
hanging-drop vapour diffusion method at 20C. Crystallization of aminotripeptidase and a derivative carrying a C-terminal His tag. Native peptidase diffracts to 2.9 A, His-tag peptidase T diffracts to 2.6 A, the selenomethionine derivative of the enzyme does not yield good crystals. His-tag derivatives not only facilitates protein purification but can also result in crystals of improved quality
-
selenomethionine His-tagged peptidase T, determination of structure by multiple wavelength anomalous dispersion methodology and refined to 2.4 A resolution
-
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37
-
slow inactivation above
45
15 min, 55% loss of activity
55
-
melting temperature
60 - 70
-
inactivation above
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
bovine serum albumin stabilizes
-
denaturation by guanidine hydrochloride at room temperature. Zn2+ makes the enzyme become much more refractory to the denaturing effect of guanidine hydrochloride than Co2+
-
very unstable during purification
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20C, 0.01 M potassium phosphate buffer, pH 7.0
-
-20C, 100 mM potassium phosphate buffer, pH 7.6, 0.05 M dithiothreitol, 50% glycerol
-
-20C, bovine serum albumin, 5 mg/ml, 6 months
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
3 forms
-
ammonium precipitation and Q-Sepharose column
-
DE-52 column chromatography and Resource Q column chromatography
-
DEAE-Toyopearl column chromatography and Toyopearl HW65C column chromatography
-
partial
peptidases a and b
-
Superose 6 gel filtration
-
Toyopearl HW65C column chromatography and DEAE-Toyopearl column chromatography
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in COS-7 cells
-
expressed in Escherichia coli M15 cells
-
expressed in Escherichia coli strain M15
-
expressed in HEK-293 cells
-
expression in CHO cells
-
expression in Escherichia coli
-
overexpression in Escherichia coli
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C25S
-
mutant enzyme shows no loss or decrease of enzymatic activity
DELTA1-119
-
inactive mutant enzyme
DELTA1-159
-
inactive mutant enzyme
DELTA1-208
-
inactive mutant enzyme
DELTA1-66
-
mutant enzyme requires a concentration of Zn2+ ion at least ten-fold higher to reach maximal activity without significantly affecting kinetic parameters such as Km and Vmax compared to the full length tripeptidase
DELTA211-410
-
inactive mutant enzyme
DELTA271-410
-
inactive mutant enzyme
DELTA291-410
-
inactive mutant enzyme
DELTA361-410
-
inactive mutant enzyme
D276A
-
kcat/KM is 21% of wild-type value
D327A
-
kcat/KM is 6% of wild-type value
D360A
-
kcat/KM is 3% of wild-type value
E272A
-
kcat/KM is 3% of wild-type value
S475L
-
kcat/KM is 0.4% of wild-type value
E205A
-
inactive
E205Q
-
inactive
E636A
-
reduced activity
S603A
-
inactive
Show AA Sequence (6520 entries)
Longer loading times are possible. Please use the Sequence Search for a specific query.