BRENDA - Enzyme Database show

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
EC Number
Inhibitors
Commentary
Organism
Structure
3.4.24.30
3,4,5,6-Tetrachloro-1,2-benzoquinone
i.e. o-chloranil, 0.003 mM, most potent inactivator
Enterococcus faecalis
3.4.24.30
Diethylpyrocarbonate
partially reversed by hydroxylamine (not)
Enterococcus faecalis
3.4.24.30
ethanol
-
Enterococcus faecalis
3.4.24.30
High MW factor of rat inflammatory exudate
strong, MW: 720000-750000
Enterococcus faecalis
3.4.24.30
L-Leucine hydroxamate
-
Enterococcus faecalis
3.4.24.30
methanol
about 15% as effective as n-butanol
Enterococcus faecalis
3.4.24.30
additional information
no inhibition by 2,3-butanedinone; no inhibition by PMSF, phenylglyoxal
Enterococcus faecalis
3.4.24.30
n-butanol
-
Enterococcus faecalis
3.4.24.30
N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline
5 mM
Enterococcus faecalis
3.4.24.30
phosphoramidon
i.e. N-(alpha-L-rhamnopyranosyloxy-hydroxyphosphinyl)-L-Leu-L-Trp
Enterococcus faecalis
3.4.24.30
Propanol
-
Enterococcus faecalis
3.4.24.30
Tetranitromethane
5 mM, pH-dependent
Enterococcus faecalis
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
3.4.24.30
0.02
-
substrate P
scissile bond Phe7-Phe8
Enterococcus faecalis
3.4.24.30
0.049
-
substrate P
scissile bond Gln6-Phe7
Enterococcus faecalis
3.4.24.30
0.0725
-
Insulin B-chain
scissile bond Phe24-Phe25
Enterococcus faecalis
3.4.24.30
0.12
-
Substance P
scissile bond Gly9-Leu10
Enterococcus faecalis
Localization
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
3.4.24.30
extracellular
-
Enterococcus faecalis
-
-
3.4.24.30
soluble
-
Enterococcus faecalis
-
-
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
3.4.24.30
Zinc
requirement, metalloendopeptidase
Enterococcus faecalis
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
3.4.24.30
additional information
-
amino acid composition shows similarity to Staphylococcus aureus metalloendopeptidase
Enterococcus faecalis
3.4.24.30
29500
-
Streptococcus faecalis, FPLC gel filtration
Enterococcus faecalis
3.4.24.30
32000
-
Streptococcus faecalis, gel filtration
Enterococcus faecalis
3.4.24.30
33000
-
1 * 33000, Streptococcus faecalis, SDS-PAGE
Enterococcus faecalis
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
3.4.24.30
Enterococcus faecalis
-
var. liquefaciens, strain OG1-10
-
Purification (Commentary)
EC Number
Commentary
Organism
3.4.24.30
-
Enterococcus faecalis
Source Tissue
EC Number
Source Tissue
Commentary
Organism
Textmining
3.4.24.30
culture filtrate
-
Enterococcus faecalis
-
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
3.4.24.30
additional information
-
-
Enterococcus faecalis
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
3.4.24.30
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
-
3.4.24.30
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
-
3.4.24.30
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
-
3.4.24.30
Azoalbumin + H2O
bovine albumin, fraction V
31192
Enterococcus faecalis
?
-
-
-
-
3.4.24.30
azocasein + H2O
-
31192
Enterococcus faecalis
?
-
-
-
-
3.4.24.30
Azocoll + H2O
general protease substrate
31192
Enterococcus faecalis
?
-
-
-
-
3.4.24.30
beta-Lipotropin
cleavage site: Gly63-Phe64, poor substrate
31192
Enterococcus faecalis
?
-
-
-
-
3.4.24.30
Bovine casein-yellow + H2O
-
31192
Enterococcus faecalis
?
-
-
-
-
3.4.24.30
Bovine fibrinogen + H2O
plasminogen-free or fraction I
31192
Enterococcus faecalis
?
-
-
-
-
3.4.24.30
Bovine hemoglobin + H2O
-
31192
Enterococcus faecalis
?
-
-
-
-
3.4.24.30
Bovine pancreas insulin + H2O
poor substrate
31192
Enterococcus faecalis
?
-
-
-
-
3.4.24.30
bradykinin + H2O
-
31192
Enterococcus faecalis
?
-
-
-
-
3.4.24.30
Cholecystokinin + H2O
cleavage site: Gly29-Trp30
31192
Enterococcus faecalis
?
-
-
-
-
3.4.24.30
Collagen + H2O
high or low viscose polypetides from collagen
31192
Enterococcus faecalis
?
-
-
-
-
3.4.24.30
Gelatin + H2O
-
31192
Enterococcus faecalis
?
-
-
-
-
3.4.24.30
Glucagon + H2O
cleavage site: Trp25-Leu26
31192
Enterococcus faecalis
?
-
-
-
-
3.4.24.30
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
-
3.4.24.30
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
?
-
-
-
-
3.4.24.30
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
-
3.4.24.30
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
-
3.4.24.30
Leucine-enkephalin + H2O
cleavage site: Gly3-Phe4, poor substrate
31192
Enterococcus faecalis
?
-
-
-
-
3.4.24.30
Lys-Bradykinin + H2O
-
31192
Enterococcus faecalis
?
-
-
-
-
3.4.24.30
additional information
strict specificity for hydrophobic amino acid at position P1', exhibits endopeptidase and thermolysin-like specificity
31192
Enterococcus faecalis
?
-
-
-
-
3.4.24.30
additional information
very poor substrates are elastin-orcein or hide powder azure
31192
Enterococcus faecalis
?
-
-
-
-
3.4.24.30
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
?
-
-
-
-
3.4.24.30
N-3-(2-Furyl)acryloyl-Gly-L-Leu amide + H2O
chromogenic substrate, cleavage site: Gly-Leu, poor substrate
31192
Enterococcus faecalis
?
-
-
-
-
3.4.24.30
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
-
3.4.24.30
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
-
3.4.24.30
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
-
3.4.24.30
[Lys1]Bradykinin + H2O
-
31192
Enterococcus faecalis
?
-
-
-
-
Subunits
EC Number
Subunits
Commentary
Organism
3.4.24.30
monomer
1 * 33000, Streptococcus faecalis, SDS-PAGE
Enterococcus faecalis
Temperature Optimum [°C]
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
3.4.24.30
37
-
assay at
Enterococcus faecalis
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
3.4.24.30
additional information
-
pI: 4.6
Enterococcus faecalis
3.4.24.30
6
8
broad, azocoll as substrate
Enterococcus faecalis
3.4.24.30
7.2
-
cleavage of Ser3-Leu4 in pheromone cAD1
Enterococcus faecalis
pH Stability
EC Number
pH Stability
pH Stability Maximum
Commentary
Organism
3.4.24.30
5.6
-
above, 17 h stable in 50 mM 2-[N-morpholino]ethanesulfonic acid buffer
Enterococcus faecalis
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
3.4.24.30
3,4,5,6-Tetrachloro-1,2-benzoquinone
i.e. o-chloranil, 0.003 mM, most potent inactivator
Enterococcus faecalis
3.4.24.30
Diethylpyrocarbonate
partially reversed by hydroxylamine (not)
Enterococcus faecalis
3.4.24.30
ethanol
-
Enterococcus faecalis
3.4.24.30
High MW factor of rat inflammatory exudate
strong, MW: 720000-750000
Enterococcus faecalis
3.4.24.30
L-Leucine hydroxamate
-
Enterococcus faecalis
3.4.24.30
methanol
about 15% as effective as n-butanol
Enterococcus faecalis
3.4.24.30
additional information
no inhibition by 2,3-butanedinone; no inhibition by PMSF, phenylglyoxal
Enterococcus faecalis
3.4.24.30
n-butanol
-
Enterococcus faecalis
3.4.24.30
N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline
5 mM
Enterococcus faecalis
3.4.24.30
phosphoramidon
i.e. N-(alpha-L-rhamnopyranosyloxy-hydroxyphosphinyl)-L-Leu-L-Trp
Enterococcus faecalis
3.4.24.30
Propanol
-
Enterococcus faecalis
3.4.24.30
Tetranitromethane
5 mM, pH-dependent
Enterococcus faecalis
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
3.4.24.30
0.02
-
substrate P
scissile bond Phe7-Phe8
Enterococcus faecalis
3.4.24.30
0.049
-
substrate P
scissile bond Gln6-Phe7
Enterococcus faecalis
3.4.24.30
0.0725
-
Insulin B-chain
scissile bond Phe24-Phe25
Enterococcus faecalis
3.4.24.30
0.12
-
Substance P
scissile bond Gly9-Leu10
Enterococcus faecalis
Localization (protein specific)
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
3.4.24.30
extracellular
-
Enterococcus faecalis
-
-
3.4.24.30
soluble
-
Enterococcus faecalis
-
-
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
3.4.24.30
Zinc
requirement, metalloendopeptidase
Enterococcus faecalis
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
3.4.24.30
additional information
-
amino acid composition shows similarity to Staphylococcus aureus metalloendopeptidase
Enterococcus faecalis
3.4.24.30
29500
-
Streptococcus faecalis, FPLC gel filtration
Enterococcus faecalis
3.4.24.30
32000
-
Streptococcus faecalis, gel filtration
Enterococcus faecalis
3.4.24.30
33000
-
1 * 33000, Streptococcus faecalis, SDS-PAGE
Enterococcus faecalis
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
3.4.24.30
-
Enterococcus faecalis
Source Tissue (protein specific)
EC Number
Source Tissue
Commentary
Organism
Textmining
3.4.24.30
culture filtrate
-
Enterococcus faecalis
-
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
3.4.24.30
additional information
-
-
Enterococcus faecalis
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
3.4.24.30
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
-
3.4.24.30
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
-
3.4.24.30
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
-
3.4.24.30
Azoalbumin + H2O
bovine albumin, fraction V
31192
Enterococcus faecalis
?
-
-
-
-
3.4.24.30
azocasein + H2O
-
31192
Enterococcus faecalis
?
-
-
-
-
3.4.24.30
Azocoll + H2O
general protease substrate
31192
Enterococcus faecalis
?
-
-
-
-
3.4.24.30
beta-Lipotropin
cleavage site: Gly63-Phe64, poor substrate
31192
Enterococcus faecalis
?
-
-
-
-
3.4.24.30
Bovine casein-yellow + H2O
-
31192
Enterococcus faecalis
?
-
-
-
-
3.4.24.30
Bovine fibrinogen + H2O
plasminogen-free or fraction I
31192
Enterococcus faecalis
?
-
-
-
-
3.4.24.30
Bovine hemoglobin + H2O
-
31192
Enterococcus faecalis
?
-
-
-
-
3.4.24.30
Bovine pancreas insulin + H2O
poor substrate
31192
Enterococcus faecalis
?
-
-
-
-
3.4.24.30
bradykinin + H2O
-
31192
Enterococcus faecalis
?
-
-
-
-
3.4.24.30
Cholecystokinin + H2O
cleavage site: Gly29-Trp30
31192
Enterococcus faecalis
?
-
-
-
-
3.4.24.30
Collagen + H2O
high or low viscose polypetides from collagen
31192
Enterococcus faecalis
?
-
-
-
-
3.4.24.30
Gelatin + H2O
-
31192
Enterococcus faecalis
?
-
-
-
-
3.4.24.30
Glucagon + H2O
cleavage site: Trp25-Leu26
31192
Enterococcus faecalis
?
-
-
-
-
3.4.24.30
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
-
3.4.24.30
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
?
-
-
-
-
3.4.24.30
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
-
3.4.24.30
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
-
3.4.24.30
Leucine-enkephalin + H2O
cleavage site: Gly3-Phe4, poor substrate
31192
Enterococcus faecalis
?
-
-
-
-
3.4.24.30
Lys-Bradykinin + H2O
-
31192
Enterococcus faecalis
?
-
-
-
-
3.4.24.30
additional information
strict specificity for hydrophobic amino acid at position P1', exhibits endopeptidase and thermolysin-like specificity
31192
Enterococcus faecalis
?
-
-
-
-
3.4.24.30
additional information
very poor substrates are elastin-orcein or hide powder azure
31192
Enterococcus faecalis
?
-
-
-
-
3.4.24.30
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
?
-
-
-
-
3.4.24.30
N-3-(2-Furyl)acryloyl-Gly-L-Leu amide + H2O
chromogenic substrate, cleavage site: Gly-Leu, poor substrate
31192
Enterococcus faecalis
?
-
-
-
-
3.4.24.30
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
-
3.4.24.30
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
-
3.4.24.30
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
-
3.4.24.30
[Lys1]Bradykinin + H2O
-
31192
Enterococcus faecalis
?
-
-
-
-
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
3.4.24.30
monomer
1 * 33000, Streptococcus faecalis, SDS-PAGE
Enterococcus faecalis
Temperature Optimum [°C] (protein specific)
EC Number
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
3.4.24.30
37
-
assay at
Enterococcus faecalis
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
3.4.24.30
additional information
-
pI: 4.6
Enterococcus faecalis
3.4.24.30
6
8
broad, azocoll as substrate
Enterococcus faecalis
3.4.24.30
7.2
-
cleavage of Ser3-Leu4 in pheromone cAD1
Enterococcus faecalis
pH Stability (protein specific)
EC Number
pH Stability
pH Stability Maximum
Commentary
Organism
3.4.24.30
5.6
-
above, 17 h stable in 50 mM 2-[N-morpholino]ethanesulfonic acid buffer
Enterococcus faecalis