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Information on EC 3.4.11.24 - aminopeptidase S and Organism(s) Streptomyces griseus and UniProt Accession P80561
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3.4.11.24 aminopeptidase SSpecify your search results
Word Map
- 3.4.11.24
- aeromonas
-
calcium-activated
- proteolytica
- aminopeptidases
- phosphonate
-
double-zinc
-
aminolysis
- septatus
-
zinc-bound
-
amino-terminus
- biotechnology
- analysis
The taxonomic range for the selected organisms is: Streptomyces griseus
The expected taxonomic range for this enzyme is: Bacteria, Archaea, Eukaryota
The expected taxonomic range for this enzyme is: Bacteria, Archaea, Eukaryota
Reaction Schemes
release of an N-terminal amino acid with a preference for large hydrophobic amino-terminus residues
Synonyms
streptomyces griseus aminopeptidase, s9ap-st, transaminopeptidase, aminopeptidase s, dinuclear aminopeptidase, aminopeptidase yscco-ii, dizinc aminopeptidase, s9 aminopeptidase, streptomyces aminopeptidase, streptomyces griseus leucine aminopeptidase, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
M28.003
-
Merops-ID
SGAP
SGAP
-
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
release of an N-terminal amino acid with a preference for large hydrophobic amino-terminus residues
hydrolyses peptide bonds formed by terminal hydrophobic amino acids such as leucine, methionine, and phenylalanine
release of an N-terminal amino acid with a preference for large hydrophobic amino-terminus residues
release of an N-terminal amino acid with a preference for large hydrophobic amino-terminus residues
catalytic mechanism, Asp160, Met161, Gly201, Arg202, and Phe219 are involved, active site structure, modeling of enzyme-substrate complex
release of an N-terminal amino acid with a preference for large hydrophobic amino-terminus residues
catalytic mechanism, high preference towards large hydrophobic amino terminus residues, active site structure, Glu131 is involved in the catalytic mechanism, enzyme-substrate and enzyme-product complex modeling
-
release of an N-terminal amino acid with a preference for large hydrophobic amino-terminus residues
M161 is involved in substrate binding at the active site cleft
-
release of an N-terminal amino acid with a preference for large hydrophobic amino-terminus residues
preference for hydrophobic residues at the ultimate and the penultimate positions, D-amino acids at these positions reduce the activity, activity is not restricted by the length of substrate chains
-
release of an N-terminal amino acid with a preference for large hydrophobic amino-terminus residues
catalytic pathway and reaction mechanism, catalytic residues are Glu131 and Tyr246, Tyr246 is involved in stabilization of the reaction transition state intermediate, also residue Glu151 is involved
-
release of an N-terminal amino acid with a preference for large hydrophobic amino-terminus residues
raction mechanism, catalytic residues are Glu131 and Tyr246, residues Arg202 and Asp160 stabilize the reaction intermediate together with Glu131
-
release of an N-terminal amino acid with a preference for large hydrophobic amino-terminus residues
structure and reaction mechanism
-
release of an N-terminal amino acid with a preference for large hydrophobic amino-terminus residues
the enzyme prefers large hydrophobic aminoterminal residues, Glu131, Asp160, and Arg202 are involved in binding of the N-terminal substrate amino acid, substrate binding and reaction mechanism, tetrahedral reaction intermediate, modelling of the enzyme-substrate and enzyme-product complexes
release of an N-terminal amino acid with a preference for large hydrophobic amino-terminus residues
catalytic reaction mechanism, a single proton transfer is involved in catalysis at pH 8.0, whereas two proton transfers are implicated at pH 6.5, involvement of a zinc-bound hydroxide as the reaction nucleophile, Tyr246 polarizes the carbonyl carbon and stabilizes the transition state, enzyme-substrate interaction, overview
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
hydrolysis of peptide bond
CAS REGISTRY NUMBER
COMMENTARY
124404-20-2
-
9031-94-1
-
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
L-leucine-4-nitroanilide + H2O
L-leucine + 4-nitroaniline
-
-
?
human hemoglobin alpha-chain + H2O
?
-
hydrolysis of the first few residues to proline at the 4th position
-
?
human hemoglobin beta-chain + H2O
?
-
hydrolysis of the first few residues to proline at the 5th position
-
?
L-Leu-4-nitroanilide + H2O
L-Leu + 4-nitroaniline
-
best substrate
-
-
?
L-lysine-4-nitroanilide + H2O
L-lysine + 4-nitroaniline
-
good substrate
-
?
L-methionine-4-nitroanilide + H2O
L-methionine + 4-nitroaniline
-
very good substrate
-
?
L-phenylalanine-4-nitroanilide + H2O
L-phenylalanine + 4-nitroaniline
-
very good substrate
-
?
L-proline-4-nitroanilide + H2O
L-proline + 4-nitroaniline
-
good substrate
-
?
Leu-4-nitroanilide + H2O
Leu + 4-nitroaniline
-
assay at pH 8.0, 30°C
-
-
?
L-alanine-4-nitroanilide + H2O
L-alanine + 4-nitroaniline
-
-
-
?
L-alanine-4-nitroanilide + H2O
L-alanine + 4-nitroaniline
-
good substrate
-
?
L-leucine-4-nitroanilide + H2O
L-leucine + 4-nitroaniline
-
-
-
?
L-leucine-4-nitroanilide + H2O
L-leucine + 4-nitroaniline
-
-
-
?
L-leucine-4-nitroanilide + H2O
L-leucine + 4-nitroaniline
-
best of the amino acid 4-nitroanilide substrates
-
?
L-leucine-4-nitroanilide + H2O
L-leucine + 4-nitroaniline
-
best of the amino acid 4-nitroanilide substrates
-
?
L-valine-4-nitroanilide + H2O
L-valine + 4-nitroaniline
-
good substrate
-
?
peptide + H2O
?
-
high preference towards large hydrophobic amino terminus residues
-
?
additional information
?
-
the enzyme is active toward various peptides with different N-terminal side chains and specific toward hydrophobic ones, the enzyme also exhibits a significant activity toward the hydrolysis of the phosphodiester bis(p-nitrophenyl)phosphate, overview, active site structure involves the three auxiliary amino acid side chains of Tyr246, Glu131, and Arg202 that are involved in catalysis, modeling of substrate binding using the crystal structure of the enzyme, overview, the activity shows proton inventory and viscosity dependence, overview
-
-
?
additional information
?
-
-
the enzyme is active toward various peptides with different N-terminal side chains and specific toward hydrophobic ones, the enzyme also exhibits a significant activity toward the hydrolysis of the phosphodiester bis(p-nitrophenyl)phosphate, overview, active site structure involves the three auxiliary amino acid side chains of Tyr246, Glu131, and Arg202 that are involved in catalysis, modeling of substrate binding using the crystal structure of the enzyme, overview, the activity shows proton inventory and viscosity dependence, overview
-
-
?
additional information
?
-
-
no activity with Gly-Pro-4-nitroanilide and Ala-Pro-4-nitroanilide, no acitivity with Xaa-Pro N-terminal sequences, glycine-4-nitroanilide and acidic amino acid 4-nitroanilide are very poor substrates
-
?
additional information
?
-
-
substrates with blocked amino groups are partially hydrolyzed, poor activity with Val-Gly, Val-Leu, and Trp-Leu, Gly-Gly-Gly and D-leucine-D-leucine are no substrates
-
?
additional information
?
-
-
aminopeptidases are involved in peptides processing and degradation, and are important in uptake of nutrients, regulation, overview
-
-
?
additional information
?
-
-
no activity with L-Glu-4-nitroanilide, L-Arg-4-nitroanilide, D-Phe-4-nitroanilide, and D-Leu-4-nitroanilide, L-Ala-4-nitroanilide and L-Pro-4-nitroanilide are poor substrates
-
-
?
additional information
?
-
-
the enzyme is active on a wide variety of peptides substrates, no activity with D-Leu-D-Leu, no prolidase activity, but release of N-terminal prolyl residues
-
-
?
additional information
?
-
-
the enzyme shows broad substrate specificity preferring N-terminal Leu or Met and Phe, but is not able to hydrolyse peptide substrates bonds with formed by acidic amino acids in the P1 position or proline in the P1 or P1' position
-
-
?
additional information
?
-
-
the thermostable enzyme prefers large hydrophobic N-terminal residues in its peptide and protein substrates, a single proton transfer is involved in catalysis at pH 8.0, whereas two proton transfers are implicated at pH 6.5
-
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
additional information
?
-
-
aminopeptidases are involved in peptides processing and degradation, and are important in uptake of nutrients, regulation, overview
-
-
?
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Ca2+
Zn2+
Ca2+
Cd2+
-
metal binding modelling using titration, kinetic, and thermodynamic data, dinuclear metal enzyme, sequential binding to two metal binding sites, affected by Ca2+
Co2+
Zn2+
additional information
Zn2+
2 mol/mol of protein, tightly bound, zinc coordination amino acid residues are conserved in similar enzymes
Ca2+
-
activates, binds to the enzyme, activation level with different enzyme substrates, overview
Ca2+
-
activates, enhances stability of the enzyme, modulates the enzyme activity and affinity towards substrates and inhibitors in a structure-dependent manner, binding structure
Ca2+
-
location of binding site is in about 25 A distance from the zinc binding site, determination of structural environment
Ca2+
activates and stabilizes, influences substrate specificity, Asp173 and Asp174 are key residues in Ca2+ binding and important for enzyme activity, residues Asp3, Ile4, Asp262, and Asp266 are also involved in calcium binding but are important for enzyme stabilization, overview, binding capacity of recombinant wild-type and mutant enzymes, overview
Ca2+
-
activates the enzyme with some substrates, about 3fold with L-Leu-4-nitroanilide, and affects substrate specificity, e.g. decreases the activity with Lys-4-nitroanilide
Ca2+
-
activates, has complex effects on the enzyme, stabilizes the enzyme
Ca2+
-
affects metal binding, inhibition, and entropy of activation of the enzyme
Zn2+
-
2 mol zinc per mol of enzyme, tightly bound at the active site in a distance of 3.6 A of each other, determination of structural environment
Zn2+
the enzyme is a double-zinc aminopeptidase, metal ions are bound at the active site
Zn2+
-
the enzyme is a double-zinc aminopeptidase, the metal ions are bound at the active site, binding structure analysis
Zn2+
the enzyme is a double-zinc exopeptidase, binding structure, overview
Zn2+
-
the enzyme is a zinc-metalloenzyme containing 2 mol zinc per mol of enzyme, stabilizes the enzyme
additional information
-
substitution of the Zn2+ ion by Mn2+ or Co2+ results in altered substrate specificity, e.g. the Co2+ containing enzyme highly prefers L-alanine-4-nitroanilide
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
EDTA
complete inhibition, activity cannot be completely restored by addition of 1 mM CaCl2 alone but together with 0.0001 mM ZnCl2 by 90%
EGTA
complete inhibition, activity can be completely restored by addition of 1 mM CaCl2
phosphate
noncompetitive inhibitor with peptide substrates, the enzyme-substrate-inhibitor ternary complex is inactive, but phosphate is a competitive inhibitor toward bis(p-nitrophenyl)phosphate hydrolysis, with Ki ranging from 2.31 to 315 mM at pH 5.0-9.0
4-iodo-L-phenylalanine
-
weak inhibition, binds at the active site via the two zinc ions displacing the metal ions, binding structure analysis
EDTA
-
complete inhibition at 10 mM, reactivation by divalent metal ions
free amino acids
-
competitive inhibition, highest inhibition by L-histidine
-
L-tryptophan
-
weak inhibition, binds at the active site via the two zinc ions displacing the metal ions, binding structure analysis
additional information
-
nonchelating 1,7-phenanthroline has little effect on the activity
-
fluoride
-
noncompetitive inhibitor at pH 5.9-8.0, fluoride ion interacts equally with the free enzyme as with the enzyme-substrate complex
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
3.8
bis(4-nitrophenyl) phosphate
-
in presence of Mn-Zn-hetero-dinuclear aminopeptidase
3.9
bis(4-nitrophenyl) phosphate
-
in presence of Mn-Co-hetero-dinuclear aminopeptidase
4.5
bis(4-nitrophenyl) phosphate
-
in presence of Zn-Zn-homo-dinuclear aminopeptidase
9.5
bis(4-nitrophenyl) phosphate
-
in presence of Co-Co-homo-dinuclear aminopeptidase
9.7
bis(4-nitrophenyl) phosphate
-
in presence of Cd-Cd-homo-dinuclear aminopeptidase
10.6
bis(4-nitrophenyl) phosphate
-
in presence of Ni-Ni-homo-dinuclear aminopeptidase
11
bis(4-nitrophenyl) phosphate
-
in presence of Mn-Cd-hetero-dinuclear aminopeptidase
12.3
bis(4-nitrophenyl) phosphate
-
in presence of Mn-Mn-homo-dinuclear aminopeptidase
12.8
bis(4-nitrophenyl) phosphate
-
in presence of Mn-Ni-hetero-dinuclear aminopeptidase
0.65
L-Leu-4-nitroanilide
-
pH 8.0, 37°C, recombinant enzyme, in presence of Ca2+
2.3
L-Leu-4-nitroanilide
-
pH 8.0, 37°C, recombinant enzyme, in absence of Ca2+
2.54
L-Leu-4-nitroanilide
pH 8.0, 37°C, recombinant mutant D3A/D262G, in absence of Ca2+
2.61
L-Leu-4-nitroanilide
pH 8.0, 37°C, recombinant wild-type enzyme, in absence of Ca2+
3.47
L-Leu-4-nitroanilide
pH 8.0, 37°C, recombinant mutant E196A, in presence of Ca2+
3.79
L-Leu-4-nitroanilide
pH 8.0, 37°C, recombinant mutant E196A, in absence of Ca2+
3.91
L-Leu-4-nitroanilide
pH 8.0, 37°C, recombinant chimeric mutant, in presence of Ca2+
4.02
L-Leu-4-nitroanilide
pH 8.0, 37°C, recombinant chimeric mutant, in absence of Ca2+
4.02
L-Leu-4-nitroanilide
pH 8.0, 37°C, recombinant wild-type enzyme, in presence of Ca2+
4.07
L-Leu-4-nitroanilide
pH 8.0, 37°C, recombinant mutant D3A/D262G, in presence of Ca2+
0.67
L-leucine-4-nitroanilide
-
pH 8.0, 22°C, native aminopeptidases 1 and 2
0.72
L-leucine-4-nitroanilide
-
pH 8.0, 22°C, acetylated aminopeptidases 1 and 2
0.00229
Leu-4-nitroanilide
-
in presence of Ni-Ni-homo-dinuclear aminopeptidase
0.00242
Leu-4-nitroanilide
-
in presence of Mn-Ni-hetero-dinuclear aminopeptidase
0.093
Leu-4-nitroanilide
-
in presence of Co-Co-homo-dinuclear aminopeptidase
0.15
Leu-4-nitroanilide
-
in presence of Mn-Co-hetero-dinuclear aminopeptidase
0.19
Leu-4-nitroanilide
-
in presence of Mn-Cd-hetero-dinuclear aminopeptidase
0.213
Leu-4-nitroanilide
-
in presence of Cd-Cd-homo-dinuclear aminopeptidase
0.88
Leu-4-nitroanilide
-
in presence of Mn-Mn-homo-dinuclear aminopeptidase
3.27
Leu-4-nitroanilide
-
in presence of Zn-Zn-homo-dinuclear aminopeptidase
3.31
Leu-4-nitroanilide
-
in presence of Mn-Zn-hetero-dinuclear aminopeptidase
additional information
additional information
kinetic analysis, effects of pH, solvent isotope effects, overview
-
additional information
additional information
-
kinetic analysis, effects of pH, solvent isotope effects, overview
-
additional information
additional information
-
kinetics and thermodynamics
-
additional information
additional information
-
kinetics,enzyme-substrate interaction, overview
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.0064
bis(4-nitrophenyl) phosphate
-
in presence of Mn-Ni-hetero-dinuclear aminopeptidase
0.01
bis(4-nitrophenyl) phosphate
-
in presence of Ni-Ni-homo-dinuclear aminopeptidase
0.016
bis(4-nitrophenyl) phosphate
-
in presence of Mn-Cd-hetero-dinuclear aminopeptidase
0.043
bis(4-nitrophenyl) phosphate
-
in presence of Cd-Cd-homo-dinuclear aminopeptidase
0.081
bis(4-nitrophenyl) phosphate
-
in presence of Mn-Mn-homo-dinuclear aminopeptidase
0.087
bis(4-nitrophenyl) phosphate
-
in presence of Mn-Co-hetero-dinuclear aminopeptidase
0.1
bis(4-nitrophenyl) phosphate
-
in presence of Mn-Zn-hetero-dinuclear aminopeptidase
0.45
bis(4-nitrophenyl) phosphate
-
in presence of Zn-Zn-homo-dinuclear aminopeptidase
0.74
bis(4-nitrophenyl) phosphate
-
in presence of Co-Co-homo-dinuclear aminopeptidase
0.49
L-Leu-4-nitroanilide
pH 8.0, 37°C, recombinant mutant E196A, in presence of Ca2+
0.55
L-Leu-4-nitroanilide
pH 8.0, 37°C, recombinant mutant E196A, in absence of Ca2+
0.97
L-Leu-4-nitroanilide
pH 8.0, 37°C, recombinant wild-type enzyme, in presence of Ca2+
1.08
L-Leu-4-nitroanilide
pH 8.0, 37°C, recombinant mutant D3A/D262G, in presence of Ca2+
2.12
L-Leu-4-nitroanilide
pH 8.0, 37°C, recombinant wild-type enzyme, in absence of Ca2+
2.71
L-Leu-4-nitroanilide
pH 8.0, 37°C, recombinant mutant D3A/D262G, in absence of Ca2+
3 - 6
L-Leu-4-nitroanilide
pH 8.0, 37°C, recombinant mutant D3A/D262G, in absence of Ca2+
4.72
L-Leu-4-nitroanilide
pH 8.0, 37°C, recombinant chimeric mutant, in presence of Ca2+
5.2
L-Leu-4-nitroanilide
pH 8.0, 37°C, recombinant chimeric mutant, in absence of Ca2+
75.3
L-Leu-4-nitroanilide
-
pH 8.0, 37°C, recombinant enzyme, in absence of Ca2+
223
L-Leu-4-nitroanilide
-
pH 8.0, 37°C, recombinant enzyme, in presence of Ca2+
75
L-leucine-4-nitroanilide
-
pH 8.0, 22°C, acetylated aminopeptidases 2
116
L-leucine-4-nitroanilide
-
pH 8.0, 22°C, acetylated aminopeptidases 1
0.0433
Leu-4-nitroanilide
-
in presence of Ni-Ni-homo-dinuclear aminopeptidase
0.0505
Leu-4-nitroanilide
-
in presence of Mn-Ni-hetero-dinuclear aminopeptidase
1.5
Leu-4-nitroanilide
-
in presence of Mn-Cd-hetero-dinuclear aminopeptidase
1.68
Leu-4-nitroanilide
-
in presence of Cd-Cd-homo-dinuclear aminopeptidase
27.5
Leu-4-nitroanilide
-
in presence of Mn-Mn-homo-dinuclear aminopeptidase
37.4
Leu-4-nitroanilide
-
in presence of Mn-Co-hetero-dinuclear aminopeptidase
41
Leu-4-nitroanilide
-
in presence of Co-Co-homo-dinuclear aminopeptidase
92.8
Leu-4-nitroanilide
-
in presence of Mn-Zn-hetero-dinuclear aminopeptidase
101
Leu-4-nitroanilide
-
in presence of Zn-Zn-homo-dinuclear aminopeptidase
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.0005
bis(4-nitrophenyl) phosphate
-
in presence of Mn-Ni-hetero-dinuclear aminopeptidase
0.00094
bis(4-nitrophenyl) phosphate
-
in presence of NNi-Ni-homo-dinuclear aminopeptidase
0.00142
bis(4-nitrophenyl) phosphate
-
in presence of Mn-Cd-hetero-dinuclear aminopeptidase
0.0044
bis(4-nitrophenyl) phosphate
-
in presence of Cd-Cd-homo-dinuclear aminopeptidase
0.00467
bis(4-nitrophenyl) phosphate
-
in presence of Mn-Mn-homo-dinuclear aminopeptidase
0.022
bis(4-nitrophenyl) phosphate
-
in presence of Mn-Co-hetero-dinuclear aminopeptidase
0.027
bis(4-nitrophenyl) phosphate
-
in presence of Mn-Zn-hetero-dinuclear aminopeptidase
0.078
bis(4-nitrophenyl) phosphate
-
in presence of Co-Co-homo-dinuclear aminopeptidase
0.1
bis(4-nitrophenyl) phosphate
-
in presence of Zn-Zn-homo-dinuclear aminopeptidase
7.89
Leu-4-nitroanilide
-
in presence of Cd-Cd-homo-dinuclear aminopeptidase
7.89
Leu-4-nitroanilide
-
in presence of Mn-Cd-hetero-dinuclear aminopeptidase
18.9
Leu-4-nitroanilide
-
in presence of Ni-Ni-homo-dinuclear aminopeptidase
20.9
Leu-4-nitroanilide
-
in presence of Mn-Ni-hetero-dinuclear aminopeptidase
28
Leu-4-nitroanilide
-
in presence of Mn-Zn-hetero-dinuclear aminopeptidase
30.9
Leu-4-nitroanilide
-
in presence of Zn-Zn-homo-dinuclear aminopeptidase
31.4
Leu-4-nitroanilide
-
in presence of Mn-Mn-homo-dinuclear aminopeptidase
249
Leu-4-nitroanilide
-
in presence of Mn-Co-hetero-dinuclear aminopeptidase
441
Leu-4-nitroanilide
-
in presence of Co-Co-homo-dinuclear aminopeptidase
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
1.1
fluoride
-
substrate Leu-4-nitroanilide, in presence of Mn-Mn-homo-dinuclear aminopeptidase
17
fluoride
-
substrate Leu-4-nitroanilide, in presence of Mn-Co-hetero-dinuclear aminopeptidase
28
fluoride
-
substrate Leu-4-nitroanilide, in presence of Co-Co-homo-dinuclear aminopeptidase
70
fluoride
-
substrate Leu-4-nitroanilide, in presence of Mn-Zn-hetero-dinuclear aminopeptidase
75
fluoride
-
substrate Leu-4-nitroanilide, in presence of Mn-Ni-hetero-dinuclear aminopeptidase
82
fluoride
-
substrate Leu-4-nitroanilide, in presence of Ni-Ni-homo-dinuclear aminopeptidase
108
fluoride
-
substrate Leu-4-nitroanilide, in presence of Zn-Zn-homo-dinuclear aminopeptidase
0.0005
L-leucine chloromethyl ketone
-
pH 8.0, 30°C, in presence of Ca2+
0.0019
L-phenylalanine chloromethyl ketone
-
pH 8.0, 30°C, in presence of Ca2+
0.0053
L-phenylalanine chloromethyl ketone
-
pH 8.0, 30°C, in absence of Ca2+
additional information
additional information
inhibition kinetics
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
625.1
additional information
additional information
recombinant wild-type and mutant enzymes, overview
additional information
-
recombinant wild-type and mutant enzymes, overview
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
additional information
-
pH-dependent activity is altered by Ca2+ concentration
additional information
-
a single proton transfer is involved in catalysis at pH 8.0, whereas two proton transfers are implicated at pH 6.5
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
37
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
pI VALUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
5.2
5.2
-
recombinant enzyme, isoelectric focusing and amino acid sequence calculation
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
protease type XIV i.e. pronase E, a protease mixture form Streptomyces griseus
protease type XIV i.e. pronase E, a protease mixture form Streptomyces griseus
-
protease type XIV i.e. pronase E, a protease mixture form Streptomyces griseus
additional information
-
the enzyme is contained in the pronase enzyme mixture from strain K-1
-
pronase preparation, containing no prolidase acivity
-
protease type XIV i.e. pronase E, a protease mixture form Streptomyces griseus
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
30000
additional information
additional information
-
extracellular enzymes show a low MW of 20-30 kDa
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
monomer
monomer
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
hanging drop vapour diffusion method, 18-25 mg/ml purified enzyme in 10 mM Tris-HCl, pH 8.0, 20 mM NaCl, 1 mM CaCl2, plus an equal volume of sodium acetate buffer at pH 5.0 to 6.0, 16-20% w/v polyethylene glycol 4000, suspended over 1 ml reservoir solution of sodium acetate, pH 5.0-6.0, 16-20% PEG 4000, 3-4 weeks, X-ray diffraction structure determination at 47.2 to 1.9 A resolution and analysis
-
protein with or without bound Zn2+ or replaced with Hg2+, hanging drop vapour diffusion method, 20 mg/ml purified enzyme in 10 mM Tris-HCl, pH 8.0, 20 mM NaCl, 1 mM CaCl2, plus an equal volume of sodium acetate buffer at pH 5.0 to 6.0, 16-20% w/v polyethylene glycol 4000, suspended over 1 ml reservoir solution of sodium acetate, pH 5.0-6.0, 16-20% PEG 4000, 4-5 weeks to full size crystals, X-ray diffraction structure determination at 2.1 to 1.75 A resolution and analysis
-
purified enzyme complexed with L-methionine, L-phenylalanine, or L-leucine, hanging-drop vapour diffusion method, protein solution: 18 mg/ml protein, 10 mM Tris-HCl, pH 8.0, 20 mM NaCl, 6 mM CaCl2, 100 mM L-methionine or 200 mM L-leucine, plus equal volume of precipitant solution: 24% w/v PEG 4000, 0.1 M ammonium sulfate, equilibrated against 1 ml of reservoir precipitant solution, 3-4 days, cyrstals of enzyme complexed with L-Phe were precipitated with 0.1 M acetate buffer, pH 5.5 instead in the same procedure within 8-10 weeks, X-ray complex structure determination at 1.6 A resolution and analysis
-
purified enzyme complexed with methionine, hanging-drop vapour diffusion method, protein solution: 18 mg/ml protein, 10 mM Tris-HCl, pH 8.0, 20 mM NaCl, 6 mM CaCl2, 0.1 M methionine, plus equal volume of precipitant solution: 24% w/v PEG 4000, 0.1 M ammonium sulfate, equilibrated against 1 ml reservoir of the precipitant solution, 3-4 days, X-ray diffraction structure determination at 1.53 A high resolution and analysis
purified enzyme in complex with tryptophan or 4-iodo-L-phenylalanine, hanging drop vapour diffusion method, 18 mg/ml protein in 10 mM Tris-HCl, pH 8.0, 20 mM NaCl, 6 mM CaCl2, and 100 mM tryptophan or 2 mM 4-iodo-L-phenylalanine, equal volumes of protein and reservoir solution are mixed, the latter containing 18% w/v PEG 4000 and 0.1 M sodium acetate, pH 5.5, equilibration against 1 ml reservoir solution for 1 day, microseeding, 3-4 days, X-ray diffraction structure determination and analysis at 1.3 A resolution
-
purified native enzyme in complex with product analogous weak inhibiting amino acids phenylalanine, leucine, and methionine, hanging drop vapour diffusion method, 18 mg/ml protein in 10 mM Tris-HCl, pH 8.0, 20 mM NaCl, 6 mM CaCl2, and 100 mM L-methionine or 200 mM L-leucine, the precipitant solution contains 24% w/v PEG 4000 and 0.1 M ammonium sulfate, equilibration against 1 ml reservoir solution, microseeding, 3-4 days, with phenylalanine a protein solution containing 18 mg/ml protein, 10 mM Tris-HCl, pH 8.0, 20 mM NaCl, 6 mM CaCl2, and 100 mM Phe is mixed with a reservoir solution containing 18% w/v PEG 4000, and 0.1 M acetate buffer, pH 5.5, microseeding, 3-4 days, X-ray diffraction structure determination and analysis at 1.8 A, 1.7 A, and 1.53 A resolution, respectively, structure modelling
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
D3A/D262G
site-directed mutagenesis, the mutant shows increased activity compared to the wild-type enzyme
E131
-
site-directed mutagenesis, the mutant enzyme shows reduced activity compared to the wild-type enzyme
E131D
-
a general acid-base mutant, thermodynamic parameters for the reaction are similar to the wild-type enzyme, but the kcat of the mutant is 4fold reduced, while the activation energy is elevated compared to the wild-type enzyme
E196A
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Y246
-
site-directed mutagenesis, the mutant enzyme shows highly reduced activity compared to the wild-type enzyme
additional information
additional information
construction of chimeric enzymes from the enzymes of Streptomyces griseus and Streptomyces septatus by DNA in vivo shuffling and site-directed mutagenesis for calcium activation and binding studies, overview
additional information
-
construction of chimeric enzymes from the enzymes of Streptomyces griseus and Streptomyces septatus by DNA in vivo shuffling and site-directed mutagenesis for calcium activation and binding studies, overview
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
69
stable during heating for 5 h at 69°C as part of the purification process
70
75
-
rate constant for inactivation Kin is 0.00028/s in absence of Ca2+, and 0.000025/s in presence of Ca2+
additional information
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20°C, liquid and freeze-dried, stable for at least 8 weeks, no loss of activity
-
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
from commercial product, 2 peaks API and APII, endopeptidase is eliminated
from commercial product, purification of the acetylated enzyme derivatives by DEAE cellulose chromatography
-
recombinant enzyme from Streptomyces lividans cells by diafiltration, hydrophobic interaction chromatography, and gel filtration to homogeneity
-
recombinant from Streptomyces lividans cell culture medium, high purity, 19% recovery
-
recombinant soluble wild-type and mutant enzymes 28fold from Escherichia coli strain BL21(DE3) by heat treatment for 20 min at 50°C and anion exchange chromatography
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
DNA and amino acid sequence determination, subcloning and expression of the recombinant enzyme possessing a Asp70 and Asp184 residues in Escherichia coli strain BL21(DE3), the soluble enzyme is inducible by growth on 1 M sorbitol, optimization of the expression method
-
expression in Streptomyces lividans by insertional cloning and protoplast transformation, the expression system contains the constitutive Streptomyces fradiae aph promoter
-
gene Sgap, DNA and amino acid sequence determination and analysis, overexpression as secreted enzyme in Escherichia coli
-
gene Sgap, screening of 21 strains with strain NBRC12875 showing the highest activity
gene SGAP, standard protoplast transformation and expression in Streptomyces lividans, excretion to the medium
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
analysis
the enzyme is clinically important as a model for undestanding the structure and mechanism of action of other metallopeptidases
biotechnology
biotechnology
-
enzyme is attractive for diverse applications, e.g. the processing of recombinant DNA proteins and fusion protein production due to its heat stability, high activity, and small size
biotechnology
the enzyme is useful in many biotechnological applications e.g. in processing of recombinant DNA, proteins, and fusion protein products
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Gilboa, R.; Greenblatt, H.M.; Perach, M.; Spungin-Bialik, A.; Lessel, U.; Wohlfahrt, G.; Schomburg, D.; Blumberg, S.; Shoham, G.
Interactions of Streptomyces griseus aminopeptidase with a methionine product analogue: a structural study at 1.53 A resolution
Acta Crystallogr. Sect. D
D56
551-558
2000
Streptomyces griseus, Streptomyces griseus (P80561)
Spungin, A.; Blumberg, S.
Streptomyces griseus aminopeptidase is a calcium-activated zinc metalloprotein. Purification and properties of the enzyme
Eur. J. Biochem.
183
471-477
1989
Streptomyces griseus (P80561), Streptomyces griseus
Ben-Meir, D.; Spungin, A.; Ashkenazi, R.; Blumberg, S.
Specificity of Streptomyces griseus aminopeptidase and modulation of activity by divalent metal ion binding and substitution
Eur. J. Biochem.
212
107-112
1993
Streptomyces griseus
Maras, B.; Greenblatt, H.M.; Shoham, G.; Spungin-Bialik, A.; Blumberg, S.; Barra, D.
Aminopeptidase from Streptomyces griseus: primary structure and comparison with other zinc-containing aminopeptidases
Eur. J. Biochem.
236
843-846
1996
Streptomyces griseus (P80561)
Papir, G.; Spungin-Bialik, A.; Ben-Meir, D.; Fudim, E.; Gilboa, R.; Greenblatt, H.M.; Shoham, G.; Lessel, U.; Schomburg, D.; Ashkenazi, R.; Blumberg, S.
Inhibition of Streptomyces griseus aminopeptidase and effects of calcium ions on catalysis and binding--comparisons with the homologous enzyme Aeromonas proteolytica aminopeptidase
Eur. J. Biochem.
258
313-319
1998
Streptomyces griseus
Vosbeck, K.D.; Greenberg, B.D.; Awad, W.M., Jr.
The proteolytic enzymes of the K-1 strain of Streptomyces griseus obtained from a commercial preparation (Pronase). Specificity and immobilization of aminopeptidase
J. Biol. Chem.
250
3981-3987
1975
Streptomyces griseus
Almog, O.; Greenblatt, H.M.; Spungin, A.; Ben-Meir, D.; Blumberg, S.; Shoham, G.
Crystallization and preliminary crystallographic analysis of Streptomyces griseus aminopeptidase
J. Mol. Biol.
230
342-344
1993
Streptomyces griseus
Greenblatt, H.M.; Almog, O.; Maras, B.; Spungin-Bialik, A.; Barra, D.; Blumberg, S.; Shoham, G.
Streptomyces griseus aminopeptidase: X-ray crystallographic structure at 1.75 A resolution
J. Mol. Biol.
265
620-636
1997
Streptomyces griseus
Ni, S.X.; Cossar, D.; Man, A.; Norek, K.; Miller, D.; Kearse, C.; Tsvetnitsky, V.
Purification and characterization of recombinant Streptomyces griseus aminopeptidase
Protein Expr. Purif.
30
62-68
2003
Streptomyces griseus
Gilboa, R.; Spungin-Bialik, A.; Wohlfahrt, G.; Schomburg, D.; Blumberg, S.; Shoham, G.
Interactions of Streptomyces griseus aminopeptidase with amino acid reaction products and their implications toward a catalytic mechanism
Proteins
44
490-504
2001
Streptomyces griseus
Reiland, V.; Gilboa, R.; Spungin-Bialik, A.; Schomburg, D.; Shoham, Y.; Blumberg, S.; Shoham, G.
Binding of inhibitory aromatic amino acids to Streptomyces griseus aminopeptidase
Acta Crystallogr. Sect. D
60
1738-1746
2004
Streptomyces griseus
Jankiewicz, U.; Bielawski, W.
The properties and functions of bacterial aminopeptidases
Acta Microbiol. Pol.
52
217-231
2003
Streptomyces griseus
Arima, J.; Iwabuchi, M.; Hatanaka, T.
Gene cloning and overproduction of an aminopeptidase from Streptomyces septatus TH-2, and comparison with a calcium-activated enzyme from Streptomyces griseus
Biochem. Biophys. Res. Commun.
317
531-538
2004
Streptomyces griseus, Streptomyces septatus (Q75V72), Streptomyces septatus TH-2 (Q75V72), Streptomyces septatus TH-2
Fundoiano-Hershcovitz, Y.; Rabinovitch, L.; Langut, Y.; Reiland, V.; Shoham, G.; Shoham, Y.
Identification of the catalytic residues in the double-zinc aminopeptidase from Streptomyces griseus
FEBS Lett.
571
192-196
2004
Streptomyces griseus
Awad, W.M., Jr.
Streptomyces griseus aminopeptidase
Handbook of Proteolytic Enzymes (Barrett, A. J. ; Rowlings, N. D. ; Woessner, J. F. , eds. )
1
957-959
2004
Streptomyces griseus, Streptomyces griseus K-1
-
Arima, J.; Uesugi, Y.; Uraji, M.; Yatsushiro, S.; Tsuboi, S.; Iwabuchi, M.; Hatanaka, T.
Modulation of Streptomyces leucine aminopeptidase by calcium: identification and functional analysis of key residues in activation and stabilization by calcium
J. Biol. Chem.
281
5885-5894
2006
Streptomyces griseus (Q5WA30), Streptomyces griseus
Hasselgren, C.; Park, H.I.; Ming, L.J.
Metal ion binding and activation of Streptomyces griseus dinuclear aminopeptidase: cadmium(II) binding as a model
J. Biol. Inorg. Chem.
6
120-127
2001
Streptomyces griseus
Ercan, A.; Park, H.I.; Ming, L.-J.
A moonlighting dizinc aminopeptidase from Streptomyces griseus: mechanisms for peptide hydrolysis and the 4 x 1010-fold acceleration of the alternative phosphodiester hydrolysis
Biochemistry
45
13779-13793
2006
Streptomyces griseus (P80561), Streptomyces griseus
Hershcovitz, Y.F.; Gilboa2, R.; Reiland, V.; Shoham, G.; Shoham, Y.
Catalytic mechanism of SGAP, a double-zinc aminopeptidase from Streptomyces griseus
FEBS J.
274
3864-3876
2007
Streptomyces griseus
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