BRENDA - Enzyme Database show
show all sequences of 3.4.24.30

Purification and substrate specificity of a strongly hydrophobic extracellular metalloendopeptidase (gelatinase) from Streptococcus faecalis (strain 0G1-10)

Mäkinen, P.L.; Clewell, D.B.; An, F.; Mäkinen, K.K.; J. Biol. Chem. 264, 3325-3334 (1989)

Data extracted from this reference:

Inhibitors
Inhibitors
Commentary
Organism
Structure
3,4,5,6-Tetrachloro-1,2-benzoquinone
i.e. o-chloranil, 0.003 mM, most potent inactivator
Enterococcus faecalis
Diethylpyrocarbonate
partially reversed by hydroxylamine (not)
Enterococcus faecalis
ethanol
-
Enterococcus faecalis
High MW factor of rat inflammatory exudate
strong, MW: 720000-750000
Enterococcus faecalis
L-Leucine hydroxamate
-
Enterococcus faecalis
methanol
about 15% as effective as n-butanol
Enterococcus faecalis
additional information
no inhibition by 2,3-butanedinone; no inhibition by PMSF, phenylglyoxal
Enterococcus faecalis
n-butanol
-
Enterococcus faecalis
N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline
5 mM
Enterococcus faecalis
phosphoramidon
i.e. N-(alpha-L-rhamnopyranosyloxy-hydroxyphosphinyl)-L-Leu-L-Trp
Enterococcus faecalis
Propanol
-
Enterococcus faecalis
Tetranitromethane
5 mM, pH-dependent
Enterococcus faecalis
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.02
-
substrate P
scissile bond Phe7-Phe8
Enterococcus faecalis
0.049
-
substrate P
scissile bond Gln6-Phe7
Enterococcus faecalis
0.0725
-
Insulin B-chain
scissile bond Phe24-Phe25
Enterococcus faecalis
0.12
-
Substance P
scissile bond Gly9-Leu10
Enterococcus faecalis
Localization
Localization
Commentary
Organism
GeneOntology No.
Textmining
extracellular
-
Enterococcus faecalis
-
-
soluble
-
Enterococcus faecalis
-
-
Metals/Ions
Metals/Ions
Commentary
Organism
Structure
Zinc
requirement, metalloendopeptidase
Enterococcus faecalis
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
additional information
-
amino acid composition shows similarity to Staphylococcus aureus metalloendopeptidase
Enterococcus faecalis
29500
-
Streptococcus faecalis, FPLC gel filtration
Enterococcus faecalis
32000
-
Streptococcus faecalis, gel filtration
Enterococcus faecalis
33000
-
1 * 33000, Streptococcus faecalis, SDS-PAGE
Enterococcus faecalis
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Enterococcus faecalis
-
var. liquefaciens, strain OG1-10
-
Purification (Commentary)
Commentary
Organism
-
Enterococcus faecalis
Source Tissue
Source Tissue
Commentary
Organism
Textmining
culture filtrate
-
Enterococcus faecalis
-
Specific Activity [micromol/min/mg]
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
additional information
-
-
Enterococcus faecalis
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Ala-Leu-Ile-Leu-Thr-Leu-Val-Ser + H2O
i.e. iPDI, competitive inhibitor of Streptococcus faecalis sex pheromones
31192
Enterococcus faecalis
Ala-Leu-Ile-Leu-Thr + Leu-Val-Ser
-
31192
Enterococcus faecalis
-
Angiotensin I + H2O
i.e. Asp-Arg-Val-Tyr-Ile-His-Pro-Phe-His-Leu
31192
Enterococcus faecalis
Asp-Arg-Val-Tyr + Ile-His-Pro + Phe-His-Leu
-
31192
Enterococcus faecalis
-
Angiotensin II + H2O
i.e. Asp-Arg-Val-Tyr-Ile-His-Pro-Phe
31192
Enterococcus faecalis
Asp-Arg-Val-Tyr + Ile-His-Pro-Phe
-
31192
Enterococcus faecalis
-
Azoalbumin + H2O
bovine albumin, fraction V
31192
Enterococcus faecalis
?
-
-
-
-
azocasein + H2O
-
31192
Enterococcus faecalis
?
-
-
-
-
Azocoll + H2O
general protease substrate
31192
Enterococcus faecalis
?
-
-
-
-
beta-Lipotropin
cleavage site: Gly63-Phe64, poor substrate
31192
Enterococcus faecalis
?
-
-
-
-
Bovine casein-yellow + H2O
-
31192
Enterococcus faecalis
?
-
-
-
-
Bovine fibrinogen + H2O
plasminogen-free or fraction I
31192
Enterococcus faecalis
?
-
-
-
-
Bovine hemoglobin + H2O
-
31192
Enterococcus faecalis
?
-
-
-
-
Bovine pancreas insulin + H2O
poor substrate
31192
Enterococcus faecalis
?
-
-
-
-
bradykinin + H2O
-
31192
Enterococcus faecalis
?
-
-
-
-
Cholecystokinin + H2O
cleavage site: Gly29-Trp30
31192
Enterococcus faecalis
?
-
-
-
-
Collagen + H2O
high or low viscose polypetides from collagen
31192
Enterococcus faecalis
?
-
-
-
-
Gelatin + H2O
-
31192
Enterococcus faecalis
?
-
-
-
-
Glucagon + H2O
cleavage site: Trp25-Leu26
31192
Enterococcus faecalis
?
-
-
-
-
Insulin A-chain + H2O
cleavage at Gln15-Leu16 and Tyr14-Gln15, to a much lesser extent than B-chain
31192
Enterococcus faecalis
Fragments of insulin A-chain
peptides (Gln1-Gln15) and (Leu16-Asn21), another product formed: peptide (Gln1-Trp14)
31192
Enterococcus faecalis
-
Insulin B-chain + H2O
oxidized, best substrate, preferred cleavage at Phe24-Phe25, followed by His5-Leu6, after prolonged incubation: Ala14-Leu15 and His10-Leu11
31192
Enterococcus faecalis
?
-
-
-
-
Leu-Phe-Ser-Leu-Val-Leu-Ala-Gly + H2O
i.e. cADI, Streptococcus faecalis sex pheromone, poor substrate
31192
Enterococcus faecalis
Leu-Phe-Ser + Leu-Val-Leu-Ala-Gly
-
31192
Enterococcus faecalis
-
Leu-Phe-Val-Val-Thr-Leu-Val-Gly + H2O
i.e. iADI, competitive inhibitor of Streptococcus faecalis sex pheromones
31192
Enterococcus faecalis
Leu-Phe-Val-Val-Thr + Leu-Val-Gly
-
31192
Enterococcus faecalis
-
Leucine-enkephalin + H2O
cleavage site: Gly3-Phe4, poor substrate
31192
Enterococcus faecalis
?
-
-
-
-
Lys-Bradykinin + H2O
-
31192
Enterococcus faecalis
?
-
-
-
-
additional information
strict specificity for hydrophobic amino acid at position P1', exhibits endopeptidase and thermolysin-like specificity
31192
Enterococcus faecalis
?
-
-
-
-
additional information
very poor substrates are elastin-orcein or hide powder azure
31192
Enterococcus faecalis
?
-
-
-
-
additional information
No substrates are Ala-Ile-Phe-Ile-Ala-Ser (i.e. Streptococcus faecalis sex pheromone cAM373), aminoacyl-2-naphthylamides, Nalpha-benzoyl-DL-Arg 4-nitroanilide, phenylazobenzyloxycarbonyl-L-Pro-L-Leu-Gly-L-Pro-D-Arg, 2-furylacryloyl-L-Leu-Gly-L-Pro-L-Ala
31192
Enterococcus faecalis
?
-
-
-
-
N-3-(2-Furyl)acryloyl-Gly-L-Leu amide + H2O
chromogenic substrate, cleavage site: Gly-Leu, poor substrate
31192
Enterococcus faecalis
?
-
-
-
-
neurotensin + H2O
cleavage site: Tyr11-Ile12, poor substrate
31192
Enterococcus faecalis
fragments of neurotensin
peptides (Arg1-Gly9) and (Leu10-Met11) are the first occuring products
31192
Enterococcus faecalis
-
Phe-Leu-Val-Met-Phe-Leu-Ser-Gly + H2O
i.e. cPDI, Streptococcus faecalis sex pheromone, poor substrate
31192
Enterococcus faecalis
Phe-Leu-Val-Met-Phe + Leu-Ser-Gly
-
31192
Enterococcus faecalis
-
Substance P + H2O
i.e. Arg-Pro-Lys-Pro-Gln-Gln-Phe-Phe-Gly-Leu-Met-NH2
31192
Enterococcus faecalis
Arg-Pro-Lys-Pro-Gln-Gln + Phe + Phe-Gly + Leu-Met-NH2
-
31192
Enterococcus faecalis
-
[Lys1]Bradykinin + H2O
-
31192
Enterococcus faecalis
?
-
-
-
-
Subunits
Subunits
Commentary
Organism
monomer
1 * 33000, Streptococcus faecalis, SDS-PAGE
Enterococcus faecalis
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
37
-
assay at
Enterococcus faecalis
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
additional information
-
pI: 4.6
Enterococcus faecalis
6
8
broad, azocoll as substrate
Enterococcus faecalis
7.2
-
cleavage of Ser3-Leu4 in pheromone cAD1
Enterococcus faecalis
pH Stability
pH Stability
pH Stability Maximum
Commentary
Organism
5.6
-
above, 17 h stable in 50 mM 2-[N-morpholino]ethanesulfonic acid buffer
Enterococcus faecalis
Inhibitors (protein specific)
Inhibitors
Commentary
Organism
Structure
3,4,5,6-Tetrachloro-1,2-benzoquinone
i.e. o-chloranil, 0.003 mM, most potent inactivator
Enterococcus faecalis
Diethylpyrocarbonate
partially reversed by hydroxylamine (not)
Enterococcus faecalis
ethanol
-
Enterococcus faecalis
High MW factor of rat inflammatory exudate
strong, MW: 720000-750000
Enterococcus faecalis
L-Leucine hydroxamate
-
Enterococcus faecalis
methanol
about 15% as effective as n-butanol
Enterococcus faecalis
additional information
no inhibition by 2,3-butanedinone; no inhibition by PMSF, phenylglyoxal
Enterococcus faecalis
n-butanol
-
Enterococcus faecalis
N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline
5 mM
Enterococcus faecalis
phosphoramidon
i.e. N-(alpha-L-rhamnopyranosyloxy-hydroxyphosphinyl)-L-Leu-L-Trp
Enterococcus faecalis
Propanol
-
Enterococcus faecalis
Tetranitromethane
5 mM, pH-dependent
Enterococcus faecalis
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.02
-
substrate P
scissile bond Phe7-Phe8
Enterococcus faecalis
0.049
-
substrate P
scissile bond Gln6-Phe7
Enterococcus faecalis
0.0725
-
Insulin B-chain
scissile bond Phe24-Phe25
Enterococcus faecalis
0.12
-
Substance P
scissile bond Gly9-Leu10
Enterococcus faecalis
Localization (protein specific)
Localization
Commentary
Organism
GeneOntology No.
Textmining
extracellular
-
Enterococcus faecalis
-
-
soluble
-
Enterococcus faecalis
-
-
Metals/Ions (protein specific)
Metals/Ions
Commentary
Organism
Structure
Zinc
requirement, metalloendopeptidase
Enterococcus faecalis
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
additional information
-
amino acid composition shows similarity to Staphylococcus aureus metalloendopeptidase
Enterococcus faecalis
29500
-
Streptococcus faecalis, FPLC gel filtration
Enterococcus faecalis
32000
-
Streptococcus faecalis, gel filtration
Enterococcus faecalis
33000
-
1 * 33000, Streptococcus faecalis, SDS-PAGE
Enterococcus faecalis
Purification (Commentary) (protein specific)
Commentary
Organism
-
Enterococcus faecalis
Source Tissue (protein specific)
Source Tissue
Commentary
Organism
Textmining
culture filtrate
-
Enterococcus faecalis
-
Specific Activity [micromol/min/mg] (protein specific)
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
additional information
-
-
Enterococcus faecalis
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Ala-Leu-Ile-Leu-Thr-Leu-Val-Ser + H2O
i.e. iPDI, competitive inhibitor of Streptococcus faecalis sex pheromones
31192
Enterococcus faecalis
Ala-Leu-Ile-Leu-Thr + Leu-Val-Ser
-
31192
Enterococcus faecalis
-
Angiotensin I + H2O
i.e. Asp-Arg-Val-Tyr-Ile-His-Pro-Phe-His-Leu
31192
Enterococcus faecalis
Asp-Arg-Val-Tyr + Ile-His-Pro + Phe-His-Leu
-
31192
Enterococcus faecalis
-
Angiotensin II + H2O
i.e. Asp-Arg-Val-Tyr-Ile-His-Pro-Phe
31192
Enterococcus faecalis
Asp-Arg-Val-Tyr + Ile-His-Pro-Phe
-
31192
Enterococcus faecalis
-
Azoalbumin + H2O
bovine albumin, fraction V
31192
Enterococcus faecalis
?
-
-
-
-
azocasein + H2O
-
31192
Enterococcus faecalis
?
-
-
-
-
Azocoll + H2O
general protease substrate
31192
Enterococcus faecalis
?
-
-
-
-
beta-Lipotropin
cleavage site: Gly63-Phe64, poor substrate
31192
Enterococcus faecalis
?
-
-
-
-
Bovine casein-yellow + H2O
-
31192
Enterococcus faecalis
?
-
-
-
-
Bovine fibrinogen + H2O
plasminogen-free or fraction I
31192
Enterococcus faecalis
?
-
-
-
-
Bovine hemoglobin + H2O
-
31192
Enterococcus faecalis
?
-
-
-
-
Bovine pancreas insulin + H2O
poor substrate
31192
Enterococcus faecalis
?
-
-
-
-
bradykinin + H2O
-
31192
Enterococcus faecalis
?
-
-
-
-
Cholecystokinin + H2O
cleavage site: Gly29-Trp30
31192
Enterococcus faecalis
?
-
-
-
-
Collagen + H2O
high or low viscose polypetides from collagen
31192
Enterococcus faecalis
?
-
-
-
-
Gelatin + H2O
-
31192
Enterococcus faecalis
?
-
-
-
-
Glucagon + H2O
cleavage site: Trp25-Leu26
31192
Enterococcus faecalis
?
-
-
-
-
Insulin A-chain + H2O
cleavage at Gln15-Leu16 and Tyr14-Gln15, to a much lesser extent than B-chain
31192
Enterococcus faecalis
Fragments of insulin A-chain
peptides (Gln1-Gln15) and (Leu16-Asn21), another product formed: peptide (Gln1-Trp14)
31192
Enterococcus faecalis
-
Insulin B-chain + H2O
oxidized, best substrate, preferred cleavage at Phe24-Phe25, followed by His5-Leu6, after prolonged incubation: Ala14-Leu15 and His10-Leu11
31192
Enterococcus faecalis
?
-
-
-
-
Leu-Phe-Ser-Leu-Val-Leu-Ala-Gly + H2O
i.e. cADI, Streptococcus faecalis sex pheromone, poor substrate
31192
Enterococcus faecalis
Leu-Phe-Ser + Leu-Val-Leu-Ala-Gly
-
31192
Enterococcus faecalis
-
Leu-Phe-Val-Val-Thr-Leu-Val-Gly + H2O
i.e. iADI, competitive inhibitor of Streptococcus faecalis sex pheromones
31192
Enterococcus faecalis
Leu-Phe-Val-Val-Thr + Leu-Val-Gly
-
31192
Enterococcus faecalis
-
Leucine-enkephalin + H2O
cleavage site: Gly3-Phe4, poor substrate
31192
Enterococcus faecalis
?
-
-
-
-
Lys-Bradykinin + H2O
-
31192
Enterococcus faecalis
?
-
-
-
-
additional information
strict specificity for hydrophobic amino acid at position P1', exhibits endopeptidase and thermolysin-like specificity
31192
Enterococcus faecalis
?
-
-
-
-
additional information
very poor substrates are elastin-orcein or hide powder azure
31192
Enterococcus faecalis
?
-
-
-
-
additional information
No substrates are Ala-Ile-Phe-Ile-Ala-Ser (i.e. Streptococcus faecalis sex pheromone cAM373), aminoacyl-2-naphthylamides, Nalpha-benzoyl-DL-Arg 4-nitroanilide, phenylazobenzyloxycarbonyl-L-Pro-L-Leu-Gly-L-Pro-D-Arg, 2-furylacryloyl-L-Leu-Gly-L-Pro-L-Ala
31192
Enterococcus faecalis
?
-
-
-
-
N-3-(2-Furyl)acryloyl-Gly-L-Leu amide + H2O
chromogenic substrate, cleavage site: Gly-Leu, poor substrate
31192
Enterococcus faecalis
?
-
-
-
-
neurotensin + H2O
cleavage site: Tyr11-Ile12, poor substrate
31192
Enterococcus faecalis
fragments of neurotensin
peptides (Arg1-Gly9) and (Leu10-Met11) are the first occuring products
31192
Enterococcus faecalis
-
Phe-Leu-Val-Met-Phe-Leu-Ser-Gly + H2O
i.e. cPDI, Streptococcus faecalis sex pheromone, poor substrate
31192
Enterococcus faecalis
Phe-Leu-Val-Met-Phe + Leu-Ser-Gly
-
31192
Enterococcus faecalis
-
Substance P + H2O
i.e. Arg-Pro-Lys-Pro-Gln-Gln-Phe-Phe-Gly-Leu-Met-NH2
31192
Enterococcus faecalis
Arg-Pro-Lys-Pro-Gln-Gln + Phe + Phe-Gly + Leu-Met-NH2
-
31192
Enterococcus faecalis
-
[Lys1]Bradykinin + H2O
-
31192
Enterococcus faecalis
?
-
-
-
-
Subunits (protein specific)
Subunits
Commentary
Organism
monomer
1 * 33000, Streptococcus faecalis, SDS-PAGE
Enterococcus faecalis
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
37
-
assay at
Enterococcus faecalis
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
additional information
-
pI: 4.6
Enterococcus faecalis
6
8
broad, azocoll as substrate
Enterococcus faecalis
7.2
-
cleavage of Ser3-Leu4 in pheromone cAD1
Enterococcus faecalis
pH Stability (protein specific)
pH Stability
pH Stability Maximum
Commentary
Organism
5.6
-
above, 17 h stable in 50 mM 2-[N-morpholino]ethanesulfonic acid buffer
Enterococcus faecalis
Other publictions for EC 3.4.24.30
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
683566
Domann
Comparative genomic analysis f ...
no activity in Enterococcus faecalis, no activity in Enterococcus faecalis Symbioflor 1
Int. J. Med. Microbiol.
297
533-539
2007
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668832
Makinen
-
Coccolysin ...
Enterococcus faecalis
Handbook of Proteolytic Enzymes(Barrett,A. J. ,Rawlings,N. D. ,Woessner,J. F. ,Eds. )Academic Press
1
396-398
2004
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1
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3
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13
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13
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649358
Fernandez-Espla
Streptococcus thermophilus cel ...
Streptococcus thermophilus, Streptococcus thermophilus CNRZ 385
Appl. Environ. Microbiol.
66
4772-4778
2000
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12
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1
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1
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5
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12
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31193
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12
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30
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3
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