Information on EC 3.4.11.21 - aspartyl aminopeptidase

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

EC NUMBER
COMMENTARY hide
3.4.11.21
-
RECOMMENDED NAME
GeneOntology No.
aspartyl aminopeptidase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
release of an N-terminal aspartate or glutamate from a peptide, with a preference for aspartate
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of peptide bond
CAS REGISTRY NUMBER
COMMENTARY hide
9074-83-3
-
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
physiological function
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
angiotensin + H2O
Asp + des-Asp-angiotensin
show the reaction diagram
Angiotensin I + H2O
?
show the reaction diagram
angiotensin I + H2O
Asp + Arg-Val-Tyr-Ile-His-Pro-Phe-His-Leu
show the reaction diagram
angiotensin I + H2O
Asp + des-Asp-angiotensin I
show the reaction diagram
-
-
-
-
?
angiotensin I + H2O
L-Asp + des-Asp-angiotensin I
show the reaction diagram
-
-
-
?
Angiotensin II + H2O
?
show the reaction diagram
angiotensin II + H2O
Asp + angiotensin III
show the reaction diagram
angiotensin II antipeptide + H2O
?
show the reaction diagram
-
-
-
?
angiotensin II antipeptide + H2O
Glu + Gly-Val-Thr-Val-His-Pro-Val
show the reaction diagram
i.e. Glu-Gly-Val-Thr-Val-His-Pro-Val
-
-
?
angiotensinogen 1-14 + H2O
?
show the reaction diagram
-
-
-
?
angiotensinogen 1-14 + H2O
Asp + Arg-Val-Tyr-Ile-His-Pro-Phe-His-Leu-Leu-Val-Tyr-Ser
show the reaction diagram
i.e. Asp-Arg-Val-Tyr-Ile-His-Pro-Phe-His-Leu-Leu-Val-Tyr-Ser, low activity
-
-
?
Arg 2-naphthylamide + H2O
Arg + 2-naphthylamine
show the reaction diagram
-
-
-
?
Asp 2-naphthylamide + H2O
Asp + 2-naphthylamine
show the reaction diagram
Asp-2-naphthylamide + H2O
Asp + 2-naphthylamine
show the reaction diagram
Asp-7-amido-4-methylcoumarin + H2O
Asp + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
?
Asp-Ala + H2O
Asp + Ala
show the reaction diagram
-
-
-
?
Asp-Ala-Pro-chromogen + H2O
Asp + Ala-Pro-chromogen
show the reaction diagram
Asp-Ala-Pro-naphthylamide + H2O
Asp + Ala-Pro-naphthylamine
show the reaction diagram
-
preferring acidic amino acids favouring aspartyl residues, requires a free alpha-amino group
-
?
Asp-Ala-Pro-sulfamethoxazole + H2O
Asp + Ala-Pro-sulfamethoxazole
show the reaction diagram
-
-
-
?
Asp-Lys + H2O
Asp + Lys
show the reaction diagram
-
-
-
?
Asp-Lys-Ala-Leu + H2O
Asp + Lys-Ala-Leu
show the reaction diagram
Asp-p-nitroanilide + H2O
Asp + p-nitroaniline
show the reaction diagram
64% activity
-
-
?
Asp-Xaa-Xaa + H2O
Asp + Xaa-Xaa
show the reaction diagram
-
no activity on oligopeptides larger than tripeptides
-
r
Asp-Xaa-Xaa-Xaa + H2O
Asp + Xaa-Xaa-Xaa
show the reaction diagram
-
neutral or hydrophobic amino acids in P1 position preferred
-
?
aspartyl peptides + H2O
Asp + des-Asp-angiotensin
show the reaction diagram
Glu 2-naphthylamide + H2O
Glu + 2-naphthylamine
show the reaction diagram
Glu-7-amido-4-methylcoumarin + H2O
Glu + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
-
?
Glu-p-nitroanilide + H2O
Glu + p-nitroaniline
show the reaction diagram
100% activity
-
-
?
L-Asp-2-naphthylamide + H2O
L-Asp + 2-naphthylamine
show the reaction diagram
-
-
-
-
?
L-aspartyl-beta-naphthylamide + H2O
L-aspartic acid + beta-naphthylamine
show the reaction diagram
-
-
-
?
Lys 2-naphthylamide + H2O
Lys + 2-naphthylamine
show the reaction diagram
-
-
-
?
N-(alpha-L-aspartyl)-4-nitroanilide + H2O
L-aspartic acid + 4-nitroaniline
show the reaction diagram
N-(alpha-L-glutamyl)-2-naphthylamide + H2O
L-glutamic acid + 2-naphthylamine
show the reaction diagram
N-(alpha-L-glutamyl)-4-nitroanilide + H2O
L-glutamic acid + 4-nitroaniline
show the reaction diagram
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
angiotensin + H2O
Asp + des-Asp-angiotensin
show the reaction diagram
angiotensin II + H2O
Asp + angiotensin III
show the reaction diagram
-
aspartyl aminopeptidase and several other aminopeptidases, i.e. angiotensinases, are involved in angiotensin II and cholecystokinin, CCK, metabolism, angiotensin II is the principal effector of the renin-angiotensin-system, RAS, which induces vasoconstriction and increases sodium and water retention thereby leading to increased blood pressure
-
-
?
aspartyl peptides + H2O
Asp + des-Asp-angiotensin
show the reaction diagram
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ba2+
-
activation
Ca2+
-
4fold activation for Glu 2-naphthylamide at 0.1 mM and above
Mn2+
91% increase of activity at 0.4 mM; 91% increase of activity at 0.4 mM
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(S)-(+)-apomorphine
IC50 value for parasite growth is 0.004 mM
1,10-phenanthroline
1-amino-1,3-propanediphosphonic acid
-
-
3-amino-3-(P-methylphosphinyl)propionic acid
-
-
3-amino-3-phosphonopropionic acid
-
-
3-amino-3-[P-(2-carboxypropyl)phosphinyl]propionic acid
-
-
4-amino-4-(P-phenylphosphinyl)butyric acid
-
20% inhibition at 1.83 mM
4-amino-4-phosphonobutyric acid
-
-
4-amino-4-[P-(2-carboxypropyl)phosphinyl]butyric acid
-
-
4-[2-(acridin-9-ylamino)ethyl]benzene-1,2-diol
IC50 value for parasite growth is 0.0013 mM
5-amino-5-phosphonopentanoic acid
-
-
Cu2+
-
strongly inhibitory
dithiothreitol
-
88% inhibition at 1 mM
EGTA
-
slightly inhibitory from 2 mM to 5 mM
NaCl
about 30% of activity is retained in 20% (w/v) NaCl; about 30% of activity is retained in 20% (w/v) NaCl
o-phenanthroline
p-chloromercuribenzoate
-
-
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
bacitracin
-
activating at 0.2 mM
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.064
angiotensin I
-
0.25
angiotensin II
-
-
0.327 - 0.385
Asp-7-amido-4-methylcoumarin
2
Asp-Ala-Pro-naphthylamide, Asp-Ala-Pro-sulfamethoxazole
-
-
-
2.3
Asp-Ala-Pro-sulfamethoxazole
-
overexpressed enzyme
-
0.94
Asp-Xaa
-
overexpressed enzyme
-
0.05 - 2.8
Asp-Xaa-Xaa-Xaa
-
-
-
0.35
Glu 2-naphthylamide
-
-
-
0.135 - 0.136
Glu-7-amido-4-methylcoumarin
8.4
Glu-Ala-Pro-naphthylamide
-
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.05 - 0.354
Asp-7-amido-4-methylcoumarin
0.011 - 0.33
Glu-7-amido-4-methylcoumarin
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00339
(S)-(+)-apomorphine
pH 7.5, temperature not specified in the publication
0.0044
1-amino-1,3-propanediphosphonic acid
-
at pH 7.5
0.395
3-amino-3-(P-methylphosphinyl)propionic acid
-
at pH 7.5
0.0271
3-amino-3-phosphonopropionic acid
-
at pH 7.5
0.183
3-amino-3-[P-(2-carboxypropyl)phosphinyl]propionic acid
-
at pH 7.5
0.00034
4-amino-4-phosphonobutyric acid
-
at pH 7.5
0.0427
4-amino-4-[P-(2-carboxypropyl)phosphinyl]butyric acid
-
at pH 7.5
0.00135
4-[2-(acridin-9-ylamino)ethyl]benzene-1,2-diol
pH 7.5, temperature not specified in the publication
0.105
5-amino-5-phosphonopentanoic acid
-
at pH 7.5
0.4
Zn2+
-
after preincubation total inhibition
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.00013
-
normal healthy rats, soluble fraction of heart, APA; normal healthy rats, soluble fraction of lung, APA
0.00024
-
normal healthy rats, soluble fraction of adrenal gland, APA
0.00032
-
normal healthy rats, soluble fraction of kidney medulla, APA
0.00041
-
normal healthy rats, soluble fraction of kidney cortex, APA
0.013 - 0.058
-
-
0.07
-
-
0.272 - 0.332
-
overexpression in Escherichia coli
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.5 - 7
-
-
7.4
-
assay at
additional information
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.5 - 8.5
-
outside the range little activity measurable
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.03
-
sequence calculation
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
29 and 33 months old rats show a 50% decrease of enzymatic activity
Manually annotated by BRENDA team
-
predominantly left hemisphere
Manually annotated by BRENDA team
-
-
Manually annotated by BRENDA team
additional information
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
35000
-
native gel filtration, SDS-PAGE
47000
x * 47000, SDS-PAGE
52428
-
8 * 55000, SDS-PAGE, 8 * 52428, amino acid sequence calculation
54430
calculated from amino acid sequence; calculated from amino acid sequence
56000
monomer, determined by SDS-PAGE
57700
SDS-PAGE; SDS-PAGE
65000
-
8 * 65000, SDS-PAGE
70000
-
8 * 70000, high performance liquid chromatography
200000
gel filtration
270000
-
native gel filtration of the proteinase treated form
350000 - 400000
-
native gel filtration of detergent-treated form
440000
520000
-
SDS-PAGE
560000
-
high performance liquid chromatography
680000
gel filtration; native enzyme, dodecamer, determined using a Superose 6 column
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
dimer
-
electron microscopy analysis, 6.5 nm diameter for each monomer
dodecamer
homooctamer
-
8 * 65000, SDS-PAGE; 8 * 70000, high performance liquid chromatography
octamer
-
8 * 55000, SDS-PAGE, 8 * 52428, amino acid sequence calculation
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
side-chain modification
-
carbohydrate
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
in complex with zinc and substrate analogue aspartate-beta-hydroxamate, to 2.2 A resolution. Structure reveals a dodecameric machinery built by domain-swapped dimers, in agreement with electron microscopy data. For both Asp-Ala and Glu-Ala substrates, the Asp and Glu side chains fit into the P1 substrate pocket without steric constraints, while the main chain is modeled onto the hydroxamate group of aspartate-beta-hydroxamate in a position optimal for hydrolysis. The P1 substrate pocket is created by strand beta15 and the beta16-alpha12 and beta17-alpha13 loops, with the beta17-lpha13 loop lining the wall and restricting the dimensions of the pocket. This limited space disfavours bulky hydrophobic residues
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6 - 11
-
stable when stored for 1 h at 37C over the pH range 6 to 11
680832
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37
-
stable when stored for 1 h at 37C over the pH range 6 to 11
60 - 70
AAP is not precipitated by heat treatment for 30 min at 60C, only less than 10% of AAP protein is precipitated by heating at 70C
60
-
50% loss of activity in 20 min
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
2000fold to homogeneity, Triton X-100 solubilized microsomal fraction, DEAE-cellulose chromatography, immunoadsorbent chromatography; proteinase solubilized microsomal fraction, DEAE-cellulose chromatography, immunoadsorbent chromatography to homogeneity
-
722fold near homogeneity by conventional chromatography techniques
-
aspartyl aminopeptidase is purified from yeast cells, using ultracentrifugation, ammonium sulfate fractionating, and chromatography on a Bio-Gel column, on a Mono Q column, and a Superose 6 column; native enzyme to homogeneity by ultracentrifugation, ammonium sulfate fractionation, anion exchange chromatography, and gel filtration
DEAE-Sephacel gel filtration and TSK phenyl 5-PW gel filtration
Ni-NTA-agarose column chromatography and Superdex-200 gel filtration
-
Ni-Sepharose 6 Fast Flow column chromatography; Ni-Sepharose 6 Fast Flow column chromatography
non-fusion protein expressed in Escherichia coli, 440fold to homogeneity
-
partial, ammonium sulfate fractionation, gel filtration, ion-exchange chromatography
-
partially, subcellular fractionation
-
recombinant His6-tagged enzyme from Escherichia coli by nickel affinity chromatography to homogeneity
-
SDS-PAGE, used for immunization
-
soluble enzyme from brain to homogeneity by affinity chromatography, gel filtration, hydrophobic interaction and hydroxyapatite chromatography
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
DNA and amino acid sequence determination and analysis
-
DNA and amino acid sequence determination and analysis, expression in Escherichia coli as His6-tagged enzyme
-
expressed in Escherichia coli BL21(DE3) cells
expressed in Escherichia coli BL21(DE3), GFP- and hemagglutinin-tagged constructs
-
expressed in Escherichia coli; expressed in Escherichia coli
expressed in Sf9 insect cells
-
expression in Escherichia coli
gene YHR113W, located on chromosome VIII, DNA and amino acid sequence determination and analysis
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
H170F
-
inactive mutant enzyme
H33F
-
mutant enzyme with decreased turnover number
H349F
-
mutant enzyme with decreased turnover number
H352F
-
dramatically reduced activity, destabilization of quarternary structure and dissociation of the native 440000 Da enzyme
H359F
-
mutant enzyme with decreased turnover number
H363F
-
mutant enzyme with decreased turnover number
H440F
-
inactive mutant enzyme
H94F
-
inactive mutant enzyme
Renatured/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
solubilization of inclusion bodies by treatment with 0.6% Triton X-100, pH 10.0, 30 mM NaCl at 40C for 20 min
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
medicine
-
enzyme is not present in the urine of healthy rats, however, it is readily detected in the urine in rat models of mild and heavy proteinuria. Urinary levels correlate with the severity of proteinuria
synthesis
-
dipeptide sweeteners aspartame and alitame