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Information on EC 3.4.11.2 - membrane alanyl aminopeptidase and Organism(s) Escherichia coli and UniProt Accession P04825

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EC Tree
     3 Hydrolases
         3.4 Acting on peptide bonds (peptidases)
             3.4.11 Aminopeptidases
                3.4.11.2 membrane alanyl aminopeptidase
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Select one or more organisms in this record: ?
This record set is specific for:
Escherichia coli
UNIPROT: P04825 not found.
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The taxonomic range for the selected organisms is: Escherichia coli
The enzyme appears in selected viruses and cellular organisms
Reaction Schemes
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Release of an N-terminal amino acid, Xaa-/-Yaa- from a peptide, amide or arylamide. Xaa is preferably Ala, but may be most amino acids including Pro (slow action). When a terminal hydrophobic residue is followed by a prolyl residue, the two may be released as an intact Xaa-Pro dipeptide
Synonyms
aminopeptidase, aminopeptidase n, dipeptidase, aminopeptidase m, alanine aminopeptidase, apn/cd13, gp150, aminopeptidase-n, anpep, alanyl aminopeptidase, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
aminopeptidase N
the enzyme has unusual specificity, cleaving adjacent to the large, nonpolar amino acids Phe and Tyr but also cleaving next to the polar residues Lys and Arg
alanine aminopeptidase
-
-
-
-
alanine-specific aminopeptidase
-
-
-
-
alanyl aminopeptidase
-
-
-
-
Alpha-aminoacylpeptide hydrolase
-
-
-
-
amino-oligopeptidase
-
-
-
-
aminopeptidase M
-
-
-
-
aminopeptidase M II
-
-
-
-
aminopeptidase N
aminopeptidase, microsomal
-
-
-
-
APN1
-
-
-
-
APN2
-
-
-
-
CD13
-
-
-
-
CryIA(C) receptor
-
-
-
-
GP 130
-
-
-
-
GP150
-
-
-
-
L-alanine aminopeptidase
-
-
-
-
Leukemia antigen CD13
-
-
-
-
Lys-AP
-
-
-
-
Lysyl aminopeptidase
-
-
-
-
Membrane glycoprotein H11
-
-
-
-
Membrane protein p161
-
-
-
-
Microsomal aminopeptidase
-
-
-
-
Myeloid plasma membrane glycoprotein CD13
-
-
-
-
particle-bound aminopeptidase
-
-
-
-
peptidase N
-
-
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
Release of an N-terminal amino acid, Xaa-/-Yaa- from a peptide, amide or arylamide. Xaa is preferably Ala, but may be most amino acids including Pro (slow action). When a terminal hydrophobic residue is followed by a prolyl residue, the two may be released as an intact Xaa-Pro dipeptide
show the reaction diagram
residues E298 and Y381 located near the zinc ion, are involved in peptide cleavage. Residue M260 functions as a cushion to accept substrates with different N-terminal resiude sizes
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis
-
-
PATHWAY SOURCE
PATHWAYS
CAS REGISTRY NUMBER
COMMENTARY hide
9054-63-1
-
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
L-Ala-L-Phe + H2O
L-Ala + L-Phe
show the reaction diagram
-
-
-
?
L-Arg-L-Phe + H2O
L-Arg + L-Phe
show the reaction diagram
-
-
-
?
L-Lys-L-Phe + H2O
L-Lys + L-Phe
show the reaction diagram
-
-
-
?
L-Phe-L-Phe + H2O
L-Phe + L-Phe
show the reaction diagram
-
-
-
?
L-Tyr-L-Phe + H2O
L-Tyr + L-Phe
show the reaction diagram
-
-
-
?
Ala-2-naphthylamide + H2O
Ala + 2-naphthylamine
show the reaction diagram
-
-
-
?
Ala-p-nitroanilide + H2O
Ala + p-nitroaniline
show the reaction diagram
aminoacyl peptide
?
show the reaction diagram
-
-
-
-
?
casein + H2O
?
show the reaction diagram
-
assay at pH 8.0, 37°C
-
-
?
dipeptides + H2O
amino acids
show the reaction diagram
-
not: Leu-Pro
-
?
Gly-2-naphthylamide + H2O
Gly + 2-naphthylamine
show the reaction diagram
-
-
-
?
Gly-7-amido-4-methylcoumarin + H2O
7-amino-4-methylcoumarin + Gly
show the reaction diagram
-
assay at pH 8.0, 37°C
-
-
?
L-Ala-4-nitroanilide + H2O
L-Ala + 4-nitroaniline
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, 37°C
-
-
?
L-Arg-4-nitroanilide + H2O
L-Arg + 4-nitroaniline
show the reaction diagram
-
-
-
-
?
L-Asp-7-amido-4-methylcoumarin + H2O
7-amino-4-methylcoumarin + L-Asp
show the reaction diagram
-
assay at pH 8.0, 37°C
-
-
?
L-Gln-7-amido-4-methylcoumarin + H2O
7-amino-4-methylcoumarin + L-Gln
show the reaction diagram
-
assay at pH 8.0, 37°C
-
-
?
L-Leu-4-nitroanilide + H2O
L-Leu + 4-nitroaniline
show the reaction diagram
-
-
-
-
?
L-Leu-7-amido-4-methylcoumarin + H2O
7-amino-4-methylcoumarin + L-Leu
show the reaction diagram
-
assay at pH 8.0, 37°C
-
-
?
L-Lys-4-nitroanilide
4-nitroaniline + L-Lys
show the reaction diagram
-
assay at pH 8.0, 37°C
-
-
?
L-Met-7-amido-4-methylcoumarin + H2O
7-amino-4-methylcoumarin + L-Met
show the reaction diagram
-
assay at pH 8.0, 37°C
-
-
?
L-Pro-7-amido-4-methylcoumarin + H2O
7-amino-4-methylcoumarin + L-Pro
show the reaction diagram
-
assay at pH 8.0, 37°C
-
-
?
L-Thr-7-amido-4-methylcoumarin + H2O
7-amino-4-methylcoumarin + L-Thr
show the reaction diagram
-
assay at pH 8.0, 37°C
-
-
?
L-Trp-7-amido-4-methylcoumarin + H2O
7-amino-4-methylcoumarin + L-Trp
show the reaction diagram
-
assay at pH 8.0, 37°C
-
-
?
L-Tyr-7-amido-4-methylcoumarin + H2O
7-amino-4-methylcoumarin + L-Tyr
show the reaction diagram
-
assay at pH 8.0, 37°C
-
-
?
Leu-2-naphthylamide + H2O
Leu + 2-naphthylamine
show the reaction diagram
-
-
-
?
Leu-Gly + H2O
Leu + Gly
show the reaction diagram
-
-
-
?
Met-2-naphthylamide + H2O
Met + 2-naphthylamine
show the reaction diagram
-
-
-
?
oxidized insulin B chain + H2O
?
show the reaction diagram
-
assay at pH 8.0, 37°C
-
-
?
tripeptides + H2O
dipeptides + amino acids
show the reaction diagram
-
not: Gly-Pro-Ala
-
?
additional information
?
-
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
aminoacyl peptide
?
show the reaction diagram
-
-
-
-
?
additional information
?
-
-
aminopeptidase regulation mechanisms and biological functions, overview
-
-
?
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Zn2+
contains zinc
Co2+
-
reactivation after inhibition by 1,10-phenanthroline
Fe2+
-
5 mol per mol of enzyme
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(Leu)3
-
-
1,10-phenanthroline
-
-
4-chloromercuribenzoate
-
-
amastatin
-
-
arginine
-
-
bestatin
Ca2+
-
50-60% inhibition at 1 mM
Co2+
-
50-60% inhibition at 1 mM
EDTA
-
no inhibition, restores activity after inhibition by metal ions
leucine
-
-
Mg2+
-
50-60% inhibition at 1 mM
Mn2+
-
50-60% inhibition at 1 mM
pepstatin
-
-
phenylalanine
-
-
puromycin
-
-
tryptophan
-
-
tyrosine
-
-
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
-
4fold up-regulation in case of oxygen or phosphorus deficit, but not in case of nitrogen or carbon deficit
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1
Ala-2-naphthylamide
-
-
2.86
Ala-p-nitroanilide
-
-
0.74
Gly-2-naphthylamide
-
-
0.31
Gly-7-amido-4-methylcoumarin
-
-
0.38
L-Ala-4-nitroanilide
-
pH 7.0, 37°C, recombinant enzyme
0.445
L-Ala-L-Ala-L-Phe-7-amido-4-methylcoumarin
-
-
0.28
L-Arg-4-nitroanilide
-
pH 7.0, 37°C, recombinant enzyme
0.2
L-Leu-4-nitroanilide
-
pH 7.0, 37°C, recombinant enzyme
0.27
L-Leu-7-amido-4-methylcoumarin
-
-
0.382
L-Lys-4-nitroanilide
-
-
0.34
L-Met-7-amido-4-methylcoumarin
-
-
0.05
Leu-2-naphthylamide
-
-
additional information
additional information
-
steady-state kinetics
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
92
Gly-7-amido-4-methylcoumarin
-
-
354
L-Ala-4-nitroanilide
-
pH 7.0, 37°C, recombinant enzyme
53
L-Ala-L-Ala-L-Phe-7-amido-4-methylcoumarin
-
-
158
L-Arg-4-nitroanilide
-
pH 7.0, 37°C, recombinant enzyme
4.4
L-Leu-4-nitroanilide
-
pH 7.0, 37°C, recombinant enzyme
90
L-Leu-7-amido-4-methylcoumarin
-
-
290
L-Lys-4-nitroanilide
-
-
80
L-Met-7-amido-4-methylcoumarin
-
-
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
292
Gly-7-amido-4-methylcoumarin
-
-
126
L-Ala-L-Ala-L-Phe-7-amido-4-methylcoumarin
-
-
330
L-Leu-7-amido-4-methylcoumarin
-
-
1000
L-Lys-4-nitroanilide
-
-
235
L-Met-7-amido-4-methylcoumarin
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.044
using L-Phe-L-Phe as substrate, in 25 mM sodium phosphate at pH 7.5
0.046
using L-Tyr-L-Phe as substrate, in 25 mM sodium phosphate at pH 7.5
0.142
using L-Lys-L-Phe as substrate, in 25 mM sodium phosphate at pH 7.5
0.154
using L-Ala-L-Phe as substrate, in 25 mM sodium phosphate at pH 7.5
0.214
using L-Arg-L-Phe as substrate, in 25 mM sodium phosphate at pH 7.5
0.78
-
Gly-2-naphthylamide as substrate
0.83
-
value about, substrate L-Asp-7-amido-4-methylcoumarin
1.6
-
Met-2-naphthylamide as substrate
1.68
-
Ala-p-nitroanilide as substrate
11.7
-
value about, substrate L-Gln-7-amido-4-methylcoumarin
116.7
-
value about, substrate L-Leu-7-amido-4-methylcoumarin
15
-
value about, substrate L-Tyr-7-amido-4-methylcoumarin
16.7
-
value about, substrate L-Thr-7-amido-4-methylcoumarin
233.3
-
value about, substrate L-Lys-4-nitroanilide
3.3
-
value about, substrate L-Trp-7-amido-4-methylcoumarin
3.54
-
Ala-2-naphthylamide as substrate
33.3
-
value about, substrate L-Pro-7-amido-4-methylcoumarin
4.8
-
Leu-2-naphthylamide as substrate
58.3
-
value about, substrate L-Ala-L-Ala-L-Phe-7-amido-4-methylcoumarin
83.3
-
value about, substrate L-Met-7-amido-4-methylcoumarin
91.7
-
value about, substrate Gly-7-amido-4-methylcoumarin
964
-
purified recombinant enzyme, substrate L-Ala-4-nitroanilide
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7
-
assay at
8
-
assay at
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
40
-
activity was measured for 5 min
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
additional information
-
enzyme expression during whole life cycle, 4fold up-regulation in case of oxygen or phosphorus deficit, but not in case of nitrogen or carbon deficit
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
deletion mutant displays lower growth during NDHT stress. Exogenous addition of casamino acid rescues the growth of pepN deletion mutant
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
99000
x * 99000, SDS-PAGE
80000
-
gel filtration, SDS-PAGE
87000
-
1 * 87000, SDS-PAGE
90000
-
SDS-PAGE
98750
-
mass spectrometry
99000
-
gel filtration
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
x * 99000, SDS-PAGE
monomer
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
in complex with the amino acids L-arginine, L-lysine, L-phenylalanine, L-tryptophan, and L-tyrosine, hanging drop vapour diffusion method
to 2.0 A resolution, space group P3121
ligand-free protein and complex with bestatin at 1.5 and 1.6 A resolution, respectively. The enzyme is composed of an N-terminal beta-domain, a catalytic domain, a middle beta-domain, and a C-terminal alpha-domain. Residues E298 and Y381 located near the zinc ion, are involved in peptide cleavage. Residue M260 functions as a cushion to accept substrates with different N-terminal resiude sizes
-
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
E264A
catalytically inactive mutant
E298A
catalytically inactive mutant
N259D
ectopically expressed in PepN deletion mutant: N259D mutant increases the activity of PepN by 2fold
N259D/Q821E
ectopically expressed in PepN deletion mutant: N259D/Q821E double mutant decreases the activity of PepN
Q821E
ectopically expressed in PepN deletion mutant: N259D mutant increases the activity of PepN by 2fold
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.5 - 9
-
stable for 1h at 30°C
35894
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
55
-
melting temperature
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
cobalt-affinity resin chromatography
chromatography columns like DEAE cellulose, Q-sepharose and butyl toyopearl, gel filtration
-
recombinant enzyme 3.8fold from overexpressing strain BL21(DE3) by anion exchange chromatography
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli BL21 (DE3) cells
expression in Escherichia coli
gene pepN, cloning from genomic DNA of strain W3110, overexpression in strain BL21(DE3), method optimization
-
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Yoshimoto, T.; Tamesa, Y.; Gushi, K.; Murayama, N.; Tsuru, D.
An aminopeptidase N from Escherichia coli HB1010: Purification and demonstration that the enzyme possesses arylamidase and peptidase activities
Agric. Biol. Chem.
52
217-225
1988
Escherichia coli
-
Manually annotated by BRENDA team
McCaman, M.T.; Villarejo, M.R.
Structured and catalytic properties of peptidase N from Escherichia coli K-12
Arch. Biochem. Biophys.
213
384-394
1982
Escherichia coli
Manually annotated by BRENDA team
Jankiewicz, U.; Bielawski, W.
The properties and functions of bacterial aminopeptidases
Acta Microbiol. Pol.
52
217-231
2003
Escherichia coli, Latilactobacillus curvatus, Lactobacillus delbrueckii, Lactococcus lactis, Pseudomonas aeruginosa, Pseudomonas fluorescens, Streptococcus thermophilus
Manually annotated by BRENDA team
Golich, F.C.; Han, M.; Crowder, M.W.
Over-expression, purification, and characterization of aminopeptidase N from Escherichia coli
Protein Expr. Purif.
47
634-639
2006
Escherichia coli
Manually annotated by BRENDA team
Onohara, Y.; Nakajima, Y.; Ito, K.; Xu, Y.; Nakashima, K.; Ito, T.; Yoshimoto, T.
Crystallization and preliminary X-ray characterization of aminopeptidase N from Escherichia coli
Acta Crystallogr. Sect. F
62
699-701
2006
Escherichia coli (P04825), Escherichia coli
Manually annotated by BRENDA team
Ito, K.; Nakajima, Y.; Onohara, Y.; Takeo, M.; Nakashima, K.; Matsubara, F.; Ito, T.; Yoshimoto, T.
Crystal structure of aminopeptidase N (proteobacteria alanyl aminopeptidase) from Escherichia coli and conformational change of methionine 260 involved in substrate recognition
J. Biol. Chem.
281
33664-33676
2006
Escherichia coli
Manually annotated by BRENDA team
Addlagatta, A.; Gay, L.; Matthews, B.W.
Structural basis for the unusual specificity of Escherichia coli aminopeptidase N
Biochemistry
47
5303-5311
2008
Escherichia coli (P04825), Escherichia coli
Manually annotated by BRENDA team
Bhosale, M.; Pande, S.; Kumar, A.; Kairamkonda, S.; Nandi, D.
Characterization of two M17 family members in Escherichia coli, Peptidase A and Peptidase B
Biochem. Biophys. Res. Commun.
395
76-81
2010
Escherichia coli
Manually annotated by BRENDA team
Bhosale, M.; Kumar, A.; Das, M.; Bhaskarla, C.; Agarwal, V.; Nandi, D.
Catalytic activity of Peptidase N is required for adaptation of Escherichia coli to nutritional downshift and high temperature stress
Microbiol. Res.
168
56-64
2013
Escherichia coli (P04825), Escherichia coli
Manually annotated by BRENDA team