Literature summary extracted from
Yokoyama, H.; Matsui, I.
A novel thermostable membrane protease forming an operon with a stomatin homolog from the hyperthermophilic archaebacterium Pyrococcus horikoshii (2005), J. Biol. Chem., 280, 6588-6594.
Cloned(Commentary)
EC Number |
Cloned (Comment) |
Organism |
---|
3.4.21.B56 |
the N-terminal region of PH1510 (1510-N, residues 16236) is expressed in Escherichia coli |
Pyrococcus horikoshii |
Protein Variants
EC Number |
Protein Variants |
Comment |
Organism |
---|
3.4.21.B56 |
D168A |
mutant enzyme shows 3.2% activity with casein compared with the activity of the wild-type enzyme. The mutant enzyme shows a 25000 Da band instead of 45000 Da band |
Pyrococcus horikoshii |
3.4.21.B56 |
K138A |
mutant enzyme shows 1.2% activity with casein compared with the activity of the wild-type enzyme |
Pyrococcus horikoshii |
3.4.21.B56 |
S97A |
mutant enzyme shows 0.08% activity with casein compared with the activity of the wild-type enzyme |
Pyrococcus horikoshii |
3.4.21.B56 |
T62A |
mutant enzyme shows 5.5% activity with casein compared with the activity of the wild-type enzyme |
Pyrococcus horikoshii |
Inhibitors
EC Number |
Inhibitors |
Comment |
Organism |
Structure |
---|
3.4.21.B56 |
3,4-dichloroisocoumarin |
1 mM, 49% inhibition |
Pyrococcus horikoshii |
|
3.4.21.B56 |
CaCl2 |
0.1 M, 11% activity |
Pyrococcus horikoshii |
|
3.4.21.B56 |
dodecyl-beta-D-maltoside |
0.1%, 38% activity |
Pyrococcus horikoshii |
|
3.4.21.B56 |
EDTA |
5 mM, 67% inhibition |
Pyrococcus horikoshii |
|
3.4.21.B56 |
iodoacetate |
5 mM, 17% inhibition |
Pyrococcus horikoshii |
|
3.4.21.B56 |
MgCl2 |
0.1 M, 14% residual activity |
Pyrococcus horikoshii |
|
3.4.21.B56 |
MnCl2 |
0.1 M, 4% activity |
Pyrococcus horikoshii |
|
3.4.21.B56 |
NaCl |
0.1 M, 90% loss of activity |
Pyrococcus horikoshii |
|
3.4.21.B56 |
pepstatin |
0.005 mM, 21% inhibition |
Pyrococcus horikoshii |
|
3.4.21.B56 |
phenylmethylsulfonyl fluoride |
5 mM, 21% inhibition |
Pyrococcus horikoshii |
|
3.4.21.B56 |
SDS |
0.01%, 53% activity. 1% SDS, 1.2% activity |
Pyrococcus horikoshii |
|
3.4.21.B56 |
ZnCl2 |
0.1 M, 9% activity |
Pyrococcus horikoshii |
|
Localization
EC Number |
Localization |
Comment |
Organism |
GeneOntology No. |
Textmining |
---|
Molecular Weight [Da]
EC Number |
Molecular Weight [Da] |
Molecular Weight Maximum [Da] |
Comment |
Organism |
---|
3.4.21.B56 |
25000 |
- |
2 * 25000, dimer of the N-terminal fragment of PH1510 (1510-N, residues 16236), SDS-PAGE |
Pyrococcus horikoshii |
Organism
EC Number |
Organism |
UniProt |
Comment |
Textmining |
---|
3.4.21.B56 |
Pyrococcus horikoshii |
O59179 |
N-terminal region of open reading frame, PH1510 (residues 16236, designated as 1510-N) is a serine protease |
- |
Purification (Commentary)
EC Number |
Purification (Comment) |
Organism |
---|
3.4.21.B56 |
purification of the N-terminal fragment of PH1510 (1510-N, residues 16236) expressed in Escherichia coli |
Pyrococcus horikoshii |
Substrates and Products (Substrate)
EC Number |
Substrates |
Comment Substrates |
Organism |
Products |
Comment (Products) |
Rev. |
Reac. |
---|
3.4.21.B56 |
alpha-casein + H2O |
alpha-casein is not cleaved as effectively as beta-casein. 1510-N probably cleaves the substrate specifically. The cleavage sites contain many hydrophobic and aromatic residues. The 1510-N protease possibly recognizes leucine at the cleavage site |
Pyrococcus horikoshii |
? |
- |
? |
|
3.4.21.B56 |
beta-casein + H2O |
alpha-casein is not cleaved as effectively as beta-casein. 1510-N probably cleaves the substrate specifically. The cleavage sites contain many hydrophobic and aromatic residues. The 1510-N protease possibly recognizes leucine at the cleavage site |
Pyrococcus horikoshii |
? |
- |
? |
|
3.4.21.B56 |
additional information |
1510-N is a serine protease with a catalytic residue, Ser-97 |
Pyrococcus horikoshii |
? |
- |
? |
|
Subunits
EC Number |
Subunits |
Comment |
Organism |
---|
3.4.21.B56 |
dimer |
2 * 25000, dimer of the N-terminal fragment of PH1510 (1510-N, residues 16236), SDS-PAGE |
Pyrococcus horikoshii |
Synonyms
EC Number |
Synonyms |
Comment |
Organism |
---|
3.4.21.B56 |
PH1510 |
- |
Pyrococcus horikoshii |
Temperature Optimum [°C]
EC Number |
Temperature Optimum [°C] |
Temperature Optimum Maximum [°C] |
Comment |
Organism |
---|
3.4.21.B56 |
80 |
- |
assay at |
Pyrococcus horikoshii |
pH Optimum
EC Number |
pH Optimum Minimum |
pH Optimum Maximum |
Comment |
Organism |
---|
3.4.21.B56 |
5 |
6 |
- |
Pyrococcus horikoshii |