Information on EC 3.4.21.19 - glutamyl endopeptidase

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

EC NUMBER
COMMENTARY hide
3.4.21.19
-
RECOMMENDED NAME
GeneOntology No.
glutamyl endopeptidase
-
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
Preferential cleavage: Glu-/-, Asp-/-
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
condensation
-
-
formation of peptide bond
hydrolysis of peptide bond
-
-
endopeptidase
-
CAS REGISTRY NUMBER
COMMENTARY hide
137010-42-5
-
82062-91-7
formerly
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
enzyme possesses Cys at position 193 instead of highly conserved Gly, the most conserved residue of the trypsin enzyme family
-
-
Manually annotated by BRENDA team
strain OG1RF
SwissProt
Manually annotated by BRENDA team
strain OG1RF
SwissProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
strain Wood 46
-
-
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
22 kDa human growth hormone + H2O
peptide fragment 1-32 of 22 kDa human growth hormone + peptide fragment 33-191 of 22 kDa human growth hormone
show the reaction diagram
-
V8-protease digestion generates the fragment amino acids 33-191, resulting from a cleavage of the amino acids 32-33 bond
-
-
?
6-azido-4-(4-iodophenethylamino)quinazoline-labeled 49 kDa subunit of NADH-ubiquinone oxidoreductase + H2O
?
show the reaction diagram
-
proteolytic mapping of the 49 kDa subunit with V8-protease, cleavage within the sequence region Asp41-Arg63: fragment A is predicted to be the peptide Thr25-Glu248, 224 amino acids, 26.0 kDa, which is further cleaved at Glu143 and give fragment B, Thr25-Glu143, 118 amino acids, overview
-
-
?
Abz-Ala-Phe-Ala-Phe-Glu-Val-Phe-(NO2)-Tyr-Asp + H2O
Abz-Ala-Phe-Ala-Phe-Glu + Val-Phe-(NO2)-Tyr-Asp
show the reaction diagram
-
-
-
?
Ac-Ala-Ala-Asn-4-methylcoumaryl-7-amide + H2O
?
show the reaction diagram
acetyl-Asp-p-nitroanilide + H2O
acetyl-Glu + p-nitroaniline
show the reaction diagram
acetyl-Glu-4-nitrophenyl + H2O
acetyl-Glu + 4-nitrophenol
show the reaction diagram
-
-
-
?
acetyl-Glu-4-nitrophenyl ester + H2O
acetyl-Glu + 4-nitrophenol
show the reaction diagram
-
-
-
?
acetyl-Glu-p-nitroanilide + H2O
acetyl-Glu + p-nitroaniline
show the reaction diagram
Ala-Glu-4-methylcoumaryl-7-amide + H2O
?
show the reaction diagram
alpha-casein + H2O
?
show the reaction diagram
alpha1-antitrypsin + H2O
?
show the reaction diagram
-
6-bromomethyl-2-(2-furanyl)-3-hydroxychromone-labeled substrate, V8 proteinase-induced cleavage of the reactive center loop does not generate any significant change in the Cys-232 region, but inactivates the anti-PPE property of the substrate, interaction analysis, overview
-
-
?
Arg-Lys-Asp-Val-Tyr + H2O
Arg-Lys-Asp + Val-Tyr
show the reaction diagram
Arg-Lys-Glu-Val-Tyr + H2O
Arg-Lys-Glu + Val-Tyr
show the reaction diagram
azocasein + H2O
?
show the reaction diagram
benzyloxycarbonyl-Ala 2-carboxyphenylthioester + H2O
?
show the reaction diagram
benzyloxycarbonyl-Ala 2-carboxyphenylthioester + H2O
benzyloxycarbonyl-Ala + 2-carboxyphenylthiol
show the reaction diagram
-
-
-
?
benzyloxycarbonyl-Ala 3-carboxyphenylester + H2O
?
show the reaction diagram
benzyloxycarbonyl-Ala 3-carboxyphenylester + H2O
benzyloxycarbonyl-Ala + 3-carboxyphenol
show the reaction diagram
-
-
-
?
benzyloxycarbonyl-Ala 3-carboxyphenylester + H2O
benzyloxycarbonyl-Ala + 3-hydroxybenzoate
show the reaction diagram
-
-
-
?
benzyloxycarbonyl-Ala 4-carboxyphenylester + H2O
?
show the reaction diagram
benzyloxycarbonyl-Ala 4-carboxyphenylester + H2O
benzyloxycarbonyl-Ala + 4-carboxyphenol
show the reaction diagram
-
-
-
?
benzyloxycarbonyl-Ala 4-carboxyphenylester + H2O
benzyloxycarbonyl-Ala + 4-hydroxybenzoate
show the reaction diagram
-
-
-
?
benzyloxycarbonyl-Ala carboxyethylthioester + H2O
?
show the reaction diagram
benzyloxycarbonyl-Ala carboxyethylthioester + H2O
benzyloxycarbonyl-Ala + carboxyethylthiol
show the reaction diagram
-
-
-
?
benzyloxycarbonyl-Ala carboxymethylthioester + H2O
?
show the reaction diagram
benzyloxycarbonyl-Ala carboxymethylthioester + H2O
benzyloxycarbonyl-Ala + carboxymethylthiol
show the reaction diagram
-
-
-
?
benzyloxycarbonyl-Ala-Ala-Glu-methyl ester + leucine * HCl
benzyloxycarbonyl-Ala-Ala-Glu-Leu + methanol
show the reaction diagram
-
peptide synthesis
-
?
benzyloxycarbonyl-Ala-Ala-Leu-Asp-4-nitroanilide + H2O
benzyloxycarbonyl-Ala-Ala-Leu-Asp + 4-nitroaniline
show the reaction diagram
-
-
-
?
benzyloxycarbonyl-Ala-Ala-Leu-Glu-4-nitroanilide + H2O
benzyloxycarbonyl-Ala-Ala-Leu-Glu + 4-nitroaniline
show the reaction diagram
-
-
-
?
benzyloxycarbonyl-Ala-Ala-Met-Asp-4-nitroanilide + H2O
benzyloxycarbonyl-Ala-Ala-Met-Asp + 4-nitroaniline
show the reaction diagram
-
-
-
?
benzyloxycarbonyl-Ala-Ala-Met-Glu-4-nitroanilide + H2O
benzyloxycarbonyl-Ala-Ala-Met-Glu + 4-nitroaniline
show the reaction diagram
-
-
-
?
benzyloxycarbonyl-Ala-Ala-Phe-Asp-4-nitroanilide + H2O
benzyloxycarbonyl-Ala-Ala-Phe-Asp + 4-nitroaniline
show the reaction diagram
-
-
-
?
benzyloxycarbonyl-Ala-Ala-Phe-Glu-4-nitroanilide + H2O
benzyloxycarbonyl-Ala-Ala-Phe-Glu + 4-nitroaniline
show the reaction diagram
benzyloxycarbonyl-Ala-Ala-Trp-Asp-4-nitroanilide + H2O
benzyloxycarbonyl-Ala-Ala-Trp-Asp + 4-nitroaniline
show the reaction diagram
-
-
-
?
benzyloxycarbonyl-Ala-Ala-Trp-Glu-4-nitroanilide + H2O
benzyloxycarbonyl-Ala-Ala-Trp-Glu + 4-nitroaniline
show the reaction diagram
-
-
-
?
benzyloxycarbonyl-Ala-Glu-methyl ester + leucine * HCl
benzyloxycarbonyl-Ala-Glu-Leu + methanol
show the reaction diagram
-
peptide synthesis
-
?
benzyloxycarbonyl-Ala-Glu-p-nitroanilide + H2O
benzyloxycarbonyl-Ala-Glu + p-nitroaniline
show the reaction diagram
benzyloxycarbonyl-Ala-Leu-Glu-p-nitroanilide + H2O
benzyloxycarbonyl-Ala-Leu-Glu + p-nitroaniline
show the reaction diagram
benzyloxycarbonyl-Asp methyl ester + H2O
benzyloxycarbonyl-Asp + methanol
show the reaction diagram
benzyloxycarbonyl-Asp-methyl ester + Ala-Ala-D-hydroxyalanine
benzyloxycarbonyl-Asp-Ala-Ala-D-hydroxyalanine + methanol
show the reaction diagram
-
peptide synthesis, low activity at 25C, but high activity at -15C
-
?
benzyloxycarbonyl-Asp-methyl ester + Ala-Ala-hydroxyproline
benzyloxycarbonyl-Asp-Ala-Ala-hydroxyproline + methanol
show the reaction diagram
-
peptide synthesis, lower activity at 25C, but high activity at -15C
-
?
benzyloxycarbonyl-Asp-methyl ester + Ala-D-hydroxyalanine
benzyloxycarbonyl-Asp-Ala-D-hydroxyalanine + methanol
show the reaction diagram
-
peptide synthesis, very low activity at 25C, but moderate activity at -15C
-
?
benzyloxycarbonyl-Asp-methyl ester + Ala-hydroxyaspartate
benzyloxycarbonyl-Asp-Ala-hydroxyaspartate + methanol
show the reaction diagram
-
peptide synthesis, low activity at 25C, but high activity at -15C
-
?
benzyloxycarbonyl-Asp-methyl ester + Ala-hydroxyglutamate
benzyloxycarbonyl-Asp-Ala-hydroxyglutamate + methanol
show the reaction diagram
-
peptide synthesis, very low activity at 25C, but high activity at -15C
-
?
benzyloxycarbonyl-Asp-methyl ester + Ala-hydroxyproline
benzyloxycarbonyl-Asp-Ala-hydroxyproline + methanol
show the reaction diagram
-
peptide synthesis, no activity at 25C, only low activity at -15C
-
?
benzyloxycarbonyl-Asp-methyl ester + Asp-hydroxyglycine
benzyloxycarbonyl-Asp-Asp-hydroxyglycine + methanol
show the reaction diagram
-
peptide synthesis, no activity at 25C, but moderate activity at -15C
-
?
benzyloxycarbonyl-Asp-methyl ester + aspartate
benzyloxycarbonyl-Asp-Asp + methanol
show the reaction diagram
-
peptide synthesis, no activity at 25C, but high activity at -15C
-
?
benzyloxycarbonyl-Asp-methyl ester + Glu-hydroxyglycine
benzyloxycarbonyl-Asp-Glu-hydroxyglycine + methanol
show the reaction diagram
-
peptide synthesis, no activity at 25C, but moderate activity at -15C
-
?
benzyloxycarbonyl-Asp-methyl ester + glutamate
benzyloxycarbonyl-Asp-Glu + methanol
show the reaction diagram
-
peptide synthesis, no activity at 25C, but high activity at -15C
-
?
benzyloxycarbonyl-Glu methyl ester + H2O
benzyloxycarbonyl-Glu + methanol
show the reaction diagram
benzyloxycarbonyl-Glu methylthioester + H2O
benzyloxycarbonyl-Glu + methylthiol
show the reaction diagram
-
-
-
?
benzyloxycarbonyl-Glu methylthioester + H2O
benzyloxycarbonyl-L-Glu + methylthiol
show the reaction diagram
-
-
-
?
benzyloxycarbonyl-Glu-4-nitroanilide + H2O
benzyloxycarbonyl-Glu + 4-nitroaniline
show the reaction diagram
-
-
-
?
benzyloxycarbonyl-Glu-methyl ester + Ala-Ala-D-hydroxyalanine
benzyloxycarbonyl-Glu-Ala-Ala-D-hydroxyalanine + methanol
show the reaction diagram
-
peptide synthesis, low activity at 25C, but high activity at -15C
-
?
benzyloxycarbonyl-Glu-methyl ester + Ala-Ala-hydroxyproline
benzyloxycarbonyl-Glu-Ala-Ala-hydroxyproline + methanol
show the reaction diagram
-
peptide synthesis, lower activity at 25C, but high activity at -15C
-
?
benzyloxycarbonyl-Glu-methyl ester + Ala-D-hydroxyalanine
benzyloxycarbonyl-Glu-Ala-D-hydroxyalanine + methanol
show the reaction diagram
-
peptide synthesis, no activity at 25C, but moderate activity at -15C
-
?
benzyloxycarbonyl-Glu-methyl ester + Ala-hydroxyaspartate
benzyloxycarbonyl-Glu-Ala-hydroxyaspartate + methanol
show the reaction diagram
-
peptide synthesis, no activity at 25C, but high activity at -15C
-
?
benzyloxycarbonyl-Glu-methyl ester + Ala-hydroxyglutamate
benzyloxycarbonyl-Glu-Ala-hydroxyglutamate + methanol
show the reaction diagram
-
peptide synthesis, no activity at 25C, but high activity at -15C
-
?
benzyloxycarbonyl-Glu-methyl ester + Ala-hydroxyproline
benzyloxycarbonyl-Glu-Ala-hydroxyproline + methanol
show the reaction diagram
-
peptide synthesis, no activity at 25C, only low activity at -15C
-
?
benzyloxycarbonyl-Glu-methyl ester + Asp-hydroxyglycine
benzyloxycarbonyl-Glu-Asp-hydroxyglycine + methanol
show the reaction diagram
-
peptide synthesis, no activity at 25C, but moderate activity at -15C
-
?
benzyloxycarbonyl-Glu-methyl ester + aspartate
benzyloxycarbonyl-Glu-Asp + methanol
show the reaction diagram
-
peptide synthesis, no activity at 25C, but moderate activity at -15C
-
?
benzyloxycarbonyl-Glu-methyl ester + Glu-hydroxyglycine
benzyloxycarbonyl-Glu-Glu-hydroxyglycine + methanol
show the reaction diagram
-
peptide synthesis, no activity at 25C, but low activity at -15C
-
?
benzyloxycarbonyl-Glu-methyl ester + glutamate
benzyloxycarbonyl-Glu-Glu + methanol
show the reaction diagram
-
peptide synthesis, no activity at 25C, but moderate activity at -15C
-
?
benzyloxycarbonyl-Glu-methyl ester + Leu-Gly
benzyloxycarbonyl-Glu-Leu-Gly + methanol
show the reaction diagram
-
peptide synthesis
-
?
benzyloxycarbonyl-Glu-methyl ester + leucine * HCl
benzyloxycarbonyl-Glu-Leu + methanol
show the reaction diagram
-
peptide synthesis
-
?
benzyloxycarbonyl-Glu-methyl ester + Phe-Gly
benzyloxycarbonyl-Glu-Phe-Gly + methanol
show the reaction diagram
-
peptide synthesis
-
?
benzyloxycarbonyl-Glu-methyl ester + tryptophan * HCl
benzyloxycarbonyl-Glu-Trp + methanol
show the reaction diagram
-
peptide synthesis
-
?
benzyloxycarbonyl-Glu-p-nitroanilide + H2O
benzyloxycarbonyl-Glu + p-nitroaniline
show the reaction diagram
benzyloxycarbonyl-Gly-Ala-Ala-Asp-4-nitroanilide + H2O
benzyloxycarbonyl-Gly-Ala-Ala-Asp + 4-nitroaniline
show the reaction diagram
-
-
-
?
benzyloxycarbonyl-Gly-Ala-Ala-Glu-4-nitroanilide + H2O
benzyloxycarbonyl-Gly-Ala-Ala-Glu + 4-nitroaniline
show the reaction diagram
-
-
-
?
benzyloxycarbonyl-L-Ala-L-Ala-L-Leu-L-Glu-4-nitroanilide + H2O
benzyloxycarbonyl-L-Ala-L-Ala-L-Leu-L-Glu + 4-nitroaniline
show the reaction diagram
-
-
-
-
?
benzyloxycarbonyl-Leu-Glu-p-nitroanilide + H2O
benzyloxycarbonyl-Leu-Glu + p-nitroaniline
show the reaction diagram
benzyloxycarbonyl-Leu-Leu-Glu-p-nitroanilide + H2O
benzyloxycarbonyl-Leu-Leu-Glu + p-nitroaniline
show the reaction diagram
benzyloxycarbonyl-Phe-Leu-Glu-p-nitroanilide + H2O
benzyloxycarbonyl-Phe-Leu-Glu + p-nitroaniline
show the reaction diagram
benzyloxycarbonyl-Pro-Leu-Gly-S-CH2-COOH + LAFARAEAF-hydroxyglycine
benzyloxycarbonyl-PLGLAFARAEAF-hydroxyglycine + HS-CH2-COOH
show the reaction diagram
-
acylation of peptide fragment by substrate mimetic
product formation 55%
?
benzyloxycarbonyl-S-CH2-COOH + LAFARAEAF-hydroxyglycine
benzyloxycarbonyl-LAFARAEAF-hydroxyglycine + HS-CH2-COOH
show the reaction diagram
-
acylation of peptide fragment by substrate mimetic
product formation 99%
?
beta-casein + H2O
?
show the reaction diagram
-
the enzyme is highly specific and hydrolyzes peptide bonds in beta-casein predominantly on the carboxy-terminal of Glu and Asp. Pro residues are not preferred, while Met is poorly preferred at the P1' position. Glu-Met hydrolysis is less preferred in comparison to Asp-Met hydrolysis
-
-
?
beta-type parvalbumin + H2O
peptide fragments
show the reaction diagram
-
from the frog Rana catesbeiana
mass spectrometry for identification
?
bovine hemoglobin + H2O
peptide fragments
show the reaction diagram
-
in presence of SDS
peptide mapping, 2 peptide fragments are Leu76-Pro-Gly-Ala-Leu-Ser-Glu82 and Lys94-Leu-His-Val-Asp-Pro-Glu100
?
bovine insulin + H2O
bovine insulin peptide fragments
show the reaction diagram
-
-
mass spectrometric identification, detailed overview
-
?
bovine myelin basic protein + H2O
?
show the reaction diagram
carboxymethylated yeast alcohol dehydrogenase + H2O
?
show the reaction diagram
-
-
5 different peptides containing the residues 5-13. 14-19, 68-77, 102-104, 105-108
?
casein + H2O
?
show the reaction diagram
CXCR4-T140 + H2O
?
show the reaction diagram
-
T140 photolabeled CXCR4, a G-protein-coupled receptor, containing the photoreactive amino acid 4-benzoyl-L-phenylalanine, Bpa, in positions 5 or 10. V8 protease digestion of both CXCR4/125I-[Bpa5]T140 and CXCR4/125I-[Bpa10]T140 adducts generates a fragment of 6 kDa suggesting that the T140 photoanalogs labeled a fragment corresponding to Lys154-Glu179 of the receptors 4th transmembrane domain
-
-
?
equine beta-casein + H2O
equine beta-casein peptide fragments
show the reaction diagram
Gelatin + H2O
?
show the reaction diagram
-
-
-
?
Glucagon + H2O
?
show the reaction diagram
GluV8 + H2O
?
show the reaction diagram
-
degradation of the C-terminus at the Glu279-Asp280 bond is suspected to be a result from autoproteolysis
38 kDa species
-
-
glycosylated bovine insulin + H2O
glycosylated bovine insulin peptide fragments
show the reaction diagram
-
three differently glycosylated substrate forms
mass spectrometric identification, detailed overview
-
?
hemocyanin + H2O
peptide fragments
show the reaction diagram
-
hydrolysis of 2 isozymes of hemocyanin KLH1 and KLH2 from shellfish Megatura crenulata at Glu-Xaa and Asp-Xaa bonds
-
?
Hemoglobin + H2O
?
show the reaction diagram
human hemoglobin + H2O
?
show the reaction diagram
-
slpicedon consisting of a flanking region FR1, the EALER sequence, and a flanking region FR2, splicing reaction at E30-R31, facilitated by organic co-solvent-induced secondary conformation of alpha17-40 within which the sequence EALER plays a major role
-
?
human parathyroid hormone(13-34) + H2O
peptides
show the reaction diagram
insulin + H2O
?
show the reaction diagram
insulin A-chain + H2O
peptide fragments
show the reaction diagram
-
hydrolysis of Glu4-Gln5, Glu17-Asp18, and Cys11-Ser12
-
?
insulin B-chain + H2O
peptide fragments
show the reaction diagram
-
hydrolysis of Glu13-Ala14, Glu21-Arg22, Cys7-Gly8, and Cys19-Gly20
-
?
insulin-like growth factor binding protein-1 + H2O
insulin-like growth factor binding protein-1 peptide fragments
show the reaction diagram
-
from human decidual cells during gestation. The phosphorylation state influences the propensity of IGFBP-1 to proteolysis, overview. Generation of Glu C peptides by V8 protease, overview
identification of Glu C peptides, overview
-
?
L-Phe-L-Leu-L-Glu-4-nitroanilide + H2O
L-Phe-L-Leu-L-Glu + 4-nitroaniline
show the reaction diagram
Leu-Leu-Glu-4-methylcoumaryl-7-amide + H2O
?
show the reaction diagram
LHCH N-terminal peptide + H2O
?
show the reaction diagram
-
N-terminal loop and first turn in helix B of light-harvesting complex II from pea
-
-
?
Lysozyme + H2O
?
show the reaction diagram
N-acetyl-L-Glu-4-nitroanilide + H2O
N-acetyl-L-Glu + 4-nitroaniline
show the reaction diagram
-
-
-
-
?
N-benzyloxycarbonyl-alpha-glutamic-p-nitroanilide + H2O
N-benzyloxycarbonyl-alpha-glutamic acid + p-nitroaniline
show the reaction diagram
-
-
-
?
N-benzyloxycarbonyl-L-glutamyl-p-nitroanilide + H2O
N-benzyloxycarbonyl-L-glutamate + p-nitroaniline
show the reaction diagram
-
-
-
-
?
N-tert-butyloxycarbonyl-L-Glu-alpha-phenyl ester + H2O
butyloxycarbonyl-L-Glu + phenol
show the reaction diagram
-
-
-
?
Nile Red-dyed microsphere based on polypeptides PLL and PLGA as shell materials + H2O
Nile Red + degraded microsphere based on polypeptides PLL and PLGA as shell materials
show the reaction diagram
-
-
-
-
?
Oxidized insulin B-chain + H2O
?
show the reaction diagram
-
-
cleavage of bond E13-A14
-
?
p-aminobenzoyl-Ala-Phe-Ala-Phe-Glu-Val-Phe-Tyr(NO2)-Asp + H2O
p-aminobenzoyl-Ala-Phe-Ala-Phe-Glu + Val-Phe-Tyr(NO2)-Asp
show the reaction diagram
-
-
-
-
?
pore-forming alpha-toxin + H2O
pore-forming alpha-toxin peptide fragments
show the reaction diagram
prothrombin + H2O
?
show the reaction diagram
Ribonuclease + H2O
?
show the reaction diagram
succinyl-Ala-Ala-Pro-Glu-p-nitroanilide + H2O
succinyl-Ala-Ala-Pro-Glu + p-nitroaniline
show the reaction diagram
t-butyloxycarbonyl-Ala-Ala-Asp-p-nitroanilide + H2O
t-butyloxycarbonyl-Ala-Ala-Asp + p-nitroaniline
show the reaction diagram
t-butyloxycarbonyl-Ala-Ala-Glu-p-nitroanilide + H2O
t-butyloxycarbonyl-Ala-Ala-Glu + p-nitroaniline
show the reaction diagram
tert-butyloxycarbonyl-Ala-Ala-Pro-Glu-4-nitroanilide + H2O
tert-butyloxycarbonyl-Ala-Ala-Pro-Glu + 4-nitroaniline
show the reaction diagram
-
-
-
?
Z-Ala-Ala-Asn-4-methylcoumaryl-7-amide + H2O
?
show the reaction diagram
Z-Leu-Leu-Glu-4-methylcoumaryl-7-amide + H2O
?
show the reaction diagram
Z-Leu-Leu-Glu-MCA + H2O
?
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
alpha-casein + H2O
?
show the reaction diagram
-
the enzyme hydrolyses Glu(51)-Tyr(52) and Glu(50)-Glu(51) in Glu(49)-Glu(50)-Glu(51)-Tyr(52) of bovine alphs1-casein
-
-
?
Gelatin + H2O
?
show the reaction diagram
-
-
-
?
GluV8 + H2O
?
show the reaction diagram
-
degradation of the C-terminus at the Glu279-Asp280 bond is suspected to be a result from autoproteolysis
38 kDa species
-
-
prothrombin + H2O
?
show the reaction diagram
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Co2+
-
strong stimulation, up to 400%, of enzyme expression in recombinant Bacillus subtilis strain at 5 mM
Mg2+
-
stimulation of enzyme expression in recombinant Bacillus subtilis strain at 1 mM
additional information
-
enzyme is a metalloprotease
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
3,4-dichloroisocoumarin
0.3 mM, 62% inhibition of isoform -1S-SprE
4-(2-aminoethyl)benzenesulfonyl fluoride
4-chloromercuribenzoate
-
slight inhibition
antipain
-
87% inhibition
Cu2+
-
gradual inhibition of enzyme expression in recombinant Bacillus subtilis strain at 1-10 mM
diisopropyl fluorophosphate
diisopropylfluorophosphate
-
complete, irreversible inhibition
E-64
-
50% inhibition
EDTA
-
30% inhibition
Fe2+
-
gradual inhibition of enzyme expression in recombinant Bacillus subtilis strain at 1-10 mM
light
-
the activity of V8 protease is inhibited in a dose-dependent manner by exposure to laser light in the presence of light-activated antimicrobial agent methylene blue leading to degradation of the V8 protease that cannot be inhibited by the addition of a protease inhibitor
-
methylene blue
-
the activity of V8 protease is inhibited in a dose-dependent manner by exposure to laser light in the presence of light-activated antimicrobial agent methylene blue leaidng to degradation of the V8 protease that cannot be inhibited by the addition of a protease inhibitor
NaCl
1 mM, 80% inhibition of isoform -1S-SprE
Pefabloc SC
-
complete inhibition
PMSF
-
slight inhibition
tagged peptide GKH17
-
i.e. GKHKNKGKKNGKHNGWK,an antimicrobial peptide, derived from kininogen boosted through end tagging with hydrophobic oligopeptide stretches. Tagging results in enhanced killing of Gram-positive Staphylococcus aureus. Microbicidal potency increases with tag length, also in plasma, and is larger for Trp and Phe stretches than for aliphatic ones, overview
-
tagged peptide HKH17
-
i.e. HKHGHGHGKHKNKGKKN,an antimicrobial peptide, derived from kininogen boosted through end tagging with hydrophobic oligopeptide stretches. Tagging results in enhanced killing of Gram-positive Staphylococcus aureus. Microbicidal potency increases with tag length, also in plasma, and is larger for Trp and Phe stretches than for aliphatic ones, overview
-
Zn2+
-
gradual inhibition of enzyme expression in recombinant Bacillus subtilis strain at 1-10 mM
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Thermolysin
-
thermolysin v8
-
-
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
3.5 - 7.6
benzyloxycarbonyl-Ala-Ala-Glu-p-nitroanilide
4.2 - 13.3
benzyloxycarbonyl-Ala-Glu-p-nitroanilide
1.5 - 3.9
benzyloxycarbonyl-Ala-Leu-Glu-p-nitroanilide
0.5
benzyloxycarbonyl-Glu-4-nitroanilide
-
pH 8.5
1.8 - 28.4
benzyloxycarbonyl-Glu-p-nitroanilide
0.67 - 3.3
benzyloxycarbonyl-L-Ala-L-Ala-L-Leu-L-Glu-4-nitroanilide
5.5 - 16.5
benzyloxycarbonyl-Leu-Glu-p-nitroanilide
0.5 - 2.8
benzyloxycarbonyl-Leu-Leu-Glu-p-nitroanilide
1.2 - 3.9
benzyloxycarbonyl-Phe-Leu-Glu-p-nitroanilide
1.48
L-Phe-L-Leu-L-Glu-4-nitroanilide
-
pH and temperature not specified in the publication
0.3 - 4.15
Leu-Leu-Glu-4-methylcoumaryl-7-amide
0.026 - 1.027
succinyl-Ala-Ala-Pro-Glu-p-nitroanilide
11.6 - 24.5
t-butyloxycarbonyl-Ala-Ala-Asp-p-nitroanilide
4.9 - 19.7
t-butyloxycarbonyl-Ala-Ala-Glu-p-nitroanilide
additional information
additional information
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
20.8 - 40.9
benzyloxycarbonyl-Ala-Ala-Glu-p-nitroanilide
5.6 - 5.9
benzyloxycarbonyl-Ala-Glu-p-nitroanilide
60.9 - 131
benzyloxycarbonyl-Ala-Leu-Glu-p-nitroanilide
81
benzyloxycarbonyl-Glu-4-nitroanilide
Bacillus intermedius
-
pH 8.5
0.28 - 0.29
benzyloxycarbonyl-Glu-p-nitroanilide
0.027 - 16.6
benzyloxycarbonyl-L-Ala-L-Ala-L-Leu-L-Glu-4-nitroanilide
22.6 - 31.4
benzyloxycarbonyl-Leu-Glu-p-nitroanilide
30.2 - 50.8
benzyloxycarbonyl-Leu-Leu-Glu-p-nitroanilide
33.2 - 40.9
benzyloxycarbonyl-Phe-Leu-Glu-p-nitroanilide
44.4
L-Phe-L-Leu-L-Glu-4-nitroanilide
Staphylococcus aureus
-
pH and temperature not specified in the publication
6147 - 9059
Leu-Leu-Glu-4-methylcoumaryl-7-amide
16.7 - 63.3
succinyl-Ala-Ala-Pro-Glu-p-nitroanilide
0.16 - 0.54
t-butyloxycarbonyl-Ala-Ala-Asp-p-nitroanilide
11.9 - 24
t-butyloxycarbonyl-Ala-Ala-Glu-p-nitroanilide
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
30
L-Phe-L-Leu-L-Glu-4-nitroanilide
Staphylococcus aureus
-
pH and temperature not specified in the publication
41922
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
1.02
-
purified enzyme
20140
isoform -1S-SprE, pH 7.5, 37C
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4
-
and a second optimum at pH 7.9, hydrolysis of hemoglobin
7 - 7.5
isoform -1S-SprE, inversely dependent on the ionic strength of buffer
7.2
-
enzyme possesses 2 optima at pH 7.2 and pH 9.5 for reaction with casein
7.9
-
and a second optimum at pH 4.0, hydrolysis of hemoglobin
8
-
hydrolysis of benzyloxycarbonyl-Glu-p-nitroanilide
8.5
-
reaction with benzyloxycarbonyl-Glu-4-nitroanilide
9.5
-
enzyme possesses 2 optima at pH 7.2 and pH 9.5 for reaction with casein
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
50
-
in absence of Ca2+
65
-
in presence of Ca2+
additional information
-
peptide synthesis at -15C
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.5
-
isoelectric focusing
additional information
-
2 ionogenic groups
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
recombinant expression as insoluble inclusion bodies
Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
11400
-
1 * 11400, SDS-PAGE
12000
-
equilibrium sedimentation
18263
-
x * 18263, amino acid sequence
23000
-
x * 23000, SDS-PAGE
25000
-
x * 25000, SDS-PAGE
26500
-
x * 26500, SDS-PAGE
29000
-
x * 29000, SDS-PAGE
30000
-
x * 30000, SDS-PAGE
32000
-
SDS-PAGE
35000
-
SDS-PAGE, wild-type GluSW
40000
-
x * 40000, SDS-PAGE, recombinant full length protein
95000
-
x * 95000, SDS-PAGE
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
proteolytic modification
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
purified enzyme in 0.01 M Tris-HCl, pH 7.0, 2 mM CaCl2, 1.2 M potassium phosphate, with or without 3% 2-methyl-2,4-pentanediol, presence of the latter leads to a second crystal form, X-ray diffraction structure determination and analysis at 1.5-1.75 A resolution
-
7 mg/ml purified enzyme in 50 mM HEPES, pH 8.6, 50 mM KCl, 15% PEG 500 monomethylester, hanging drop method over a reservoir solution containing 0.1 M KCl, 0.1 M HEPES, and 20% PEG 5000 monomethyl ester, 293 K, X-ray diffraction structure determination and analysis at 1.9 A resolution
-
enzyme is crystallized complexed with substrate Boc-Ala-Ala-Pro-Glu-4-nitroanilide, X-ray structure determination and analysis at 2.0 A resolution
-
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.5 - 11
-
22C, 3 h, stable
81317, 81320
8
-
50C, 24 h, in presence of 5 mM Ca2+, stable
81317
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
22 - 37
-
stable, in absence of Ca2+
22
-
pH 6.5-11.0, 3 h, stable
37 - 55
-
stable, in presence of Ca2+
55
-
in absence of Ca2+, loss of 90% activity
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
Ca2+ stabilizes
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-70C, all isoforms stable for at least 6 months
4C, 10 months, no loss of activity for mutant H186T
-
4C, isoform -1S-SprE, 50% loss of activity within 4 weeks, isoform 1l-SprE, stable
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
1275fold, to homogeneity
-
full length form of C-terminally His-tagged GluSE is purified as three proenzymes equivalent to the native ones
further purification of commercial preparation
-
large scale purification as one step purification using Talon affinity chromatography
-
partial
-
phenyl Sepharose 6 column chromatography and HiTrap-CM column chromatography
-
phenyl Sepharose column chromatography, and HiTrap-CM column chromatography
-
recombinant Bacillus intermedius glutamyl endopeptidase (BIGEP-Bs) is purified from Bacillus subtilis AJ73 strain
-
recombinant fusion protein containing the mature protein sequence and the pro-sequence of the analogous enzyme from Staphylococcus epidermidis and the four point mutant enzyme in large scale using Talon affinity chromatography
-
Talon affinity column chromatography
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
different BIGEP forms (full-length precursor, precursor without signal peptide and mature part) are expressed in Escherichia coli
-
expressed as soluble His-tag fusion protein in Escherichia coli
-
expressed in Bacillus subtilis recombinant strains with mutations in the regulatory proteins Spo0A, KinA, and Ger
-
expressed in enzyme deficient strain Bacillus subtilis AJ73, sporulation and enzyme expression is strongly reduced when 1% glucose is added to the culture medium at the beginning of the growth phase, no effect of glucose addition after 20 h of cultivation period, slightly enhanced activity when glucose is added after 30 h of cultivation
-
expressed in Escherichia coli
expressed in Escherichia coli, poor expression of native full length protein, successful expression when mature protein sequence is fused with the pro-sequence of the analogous enzyme from Staphylococcus epidermidis, successful expression of double and four point mutant in Escherichia coli
-
expression in Bacillus subtilis, optimization of culture medium composition for production of recombinant enzyme, recombinant enzyme is secreted to the medium from Bacillus subtilis cells, 2-5% of the enzyme remain bound to the cell wall
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
H186T
-
no effect on enzyme specificity, 4.9fold increase in KM-value, 600fold decrease in kcat-value
DELTA1-48
-
the 38 and 40 kDa mature forms are obtained after thermolysin treatment
DELTA1-55
-
the 38 and 40 kDa mature forms are obtained after thermolysin treatment
DELTA1-60
-
the 38 and 40 kDa mature forms are obtained after thermolysin treatment
DELTA1-62
-
the 38 and 40 kDa mature forms are obtained after thermolysin treatment
DELTA1-64
-
poor expression in Escherichia coli, resulting enzyme thoroughly degraded upon thermolysin treatment, very low activity
DELTA1-65
-
poor expression in Escherichia coli, resulting enzyme thoroughly degraded upon thermolysin treatment, activity hardly detectable
E62Q/E65S
-
mutation prevent degradation of protein, slightly accelerated proliferation rate compared with wild type enzyme when expressed in Escherichia coli
E62Q/E65S/A67P/N68P
-
efficient suppression of proteolysis, strongly accelerated proliferation rate compared with wild type enzyme when expressed in Escherichia coli
G176E/Q179E/Y185W/D189P/K191E/Y192F/S194G/S195A
-
GluV8DELTAC, the C-terminal 52 residues are deleted
S237A
-
mutation introduced into the fusion protein containing the mature protein sequence and the pro-sequence of the analogous enzyme from Staphylococcus epidermidis, no proteinolytic activity
S66R
-
insertion of a trypsin degradable sequence, successful enzyme processing by trypsin instead of thermolysin, enhanced Glu-specific activity
V69A
-
mutation introduced into the fusion protein containing the mature protein sequence and the pro-sequence of the analogous enzyme from Staphylococcus epidermidis, normal processing of propeptide to mature protein, no proteinolytic activity
V69F
-
mutation introduced into the fusion protein containing the mature protein sequence and the pro-sequence of the analogous enzyme from Staphylococcus epidermidis, normal processing of propeptide to mature protein, no proteinolytic activity
V69G
-
mutation introduced into the fusion protein containing the mature protein sequence and the pro-sequence of the analogous enzyme from Staphylococcus epidermidis, normal processing of propeptide to mature protein, no proteinolytic activity
K191E/Y192F/S194G/S195A
-
GluSE-EFGA
N190H/Y192H/S169A
-
the specific activity of the mutant is 4.5fold increased from that of the wild type enzyme. The mutant potently hydrolyzes LLE-7-amido-4-methylcomarin
S66R/V69A
-
insertion of a trypsin degradable sequence at position 66, successful enzyme processing by trypsin instead of thermolysin, 4.5% of activity compared with the Val69 native form
S66R/V69F
-
insertion of a trypsin degradable sequence at position 66, successful enzyme processing by trypsin instead of thermolysin, 1.4% of activity compared with the Val69 native form
S66R/V69G
-
insertion of a trypsin degradable sequence at position 66, successful enzyme processing by trypsin instead of thermolysin, 1.1% of activity compared with the Val69 native form
S66R/V69S
-
insertion of a trypsin degradable sequence at position 66, successful enzyme processing by trypsin instead of thermolysin, 0.6% of activity compared with the Val69 native form
Y185W/D189P
-
the activity of GluSE with the two substitutions, i.e., Y185W and D189P (designated GluSE-WP), is equivalent to that of GluSE-WPEFGA
Y185W/D189P/K191E/Y192F/S194G/S195A
-
GluSE/WPEFGA
N190H/Y192H/S169A
-
the specific activity of the mutant is 4.5fold increased from that of the wild type enzyme. The mutant potently hydrolyzes LLE-7-amido-4-methylcomarin
-
H199V
-
change in the substrate preference, with a 16fold increase in the ratio of turnover number to Km-value with substrates with Phe in P1, and a 20fold increase with substrates with Glu in P1. Substitution of His199 by anything except Val completely abolishes the production of mature enzyme
H228A
-
change in substrate specificity, i.e., a slight increase in the ratio of turnover-number to Km-value for the substrate with Asp and a more than 300-fold increase in this ratio when P1 substituent is Ala
S216A
-
about 7.5fold increase in Km-value for hydrolysis of succinyl-Ala-Ala-Pro-Glu-p-nitroanilide compared to the wild-type enzyme
S216G
-
about 7.5fold increase in Km-value for hydrolysis of succinyl-Ala-Ala-Pro-Glu-p-nitroanilide compared to the wild-type enzyme
additional information
Renatured/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
recombinant expression of enzyme as insoluble inclusion bodies, solubilization in 6 M guanidine-HCl in presence of reducing agent and renaturation by fast frequent dilution method. Highest yield of refolded protein at pH 8.4, 4C. Renaturation is accompanied by gradual splitting of K12-E13 and T47-E48 bonds resulting in a 26 kDa protein
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
synthesis