Literature summary extracted from
Matsui, E.; Kawasaki, S.; Ishida, H.; Ishikawa, K.; Kosugi, Y.; Kikuchi, H.; Kawarabayashi, Y.; Matsui, I.
Thermostable flap endonuclease from the archaeon, Pyrococcus horikoshii, cleaves the replication fork-like structure endo/exonucleolytically (1999), J. Biol. Chem., 274, 18297-18309.
Cloned(Commentary)
EC Number |
Cloned (Comment) |
Organism |
---|
3.1.99.B1 |
overexpressed in Escherichia coli |
Pyrococcus horikoshii |
Inhibitors
EC Number |
Inhibitors |
Comment |
Organism |
Structure |
---|
3.1.99.B1 |
Ca2+ |
10 mM CaCl2 results in greater than 90% inhibition |
Pyrococcus horikoshii |
|
3.1.99.B1 |
EDTA |
5 mM, complete inhibition |
Pyrococcus horikoshii |
|
3.1.99.B1 |
KCl |
60 mM, the activity increases 1.3-fold, 100 mM KCl result in significant inhibition of activity |
Pyrococcus horikoshii |
|
3.1.99.B1 |
Mg2+ |
addition of 1 mM MgCl2 increases the activity 1.4-fold. 10 mM MgCl2 results in greater than 90% inhibition |
Pyrococcus horikoshii |
|
3.1.99.B1 |
Mn2+ |
addition of 0.1 mM MgCl2 increases the activity 1.5-fold. 10 mM MnCl2 results in greater than 90% inhibition |
Pyrococcus horikoshii |
|
Metals/Ions
EC Number |
Metals/Ions |
Comment |
Organism |
Structure |
---|
3.1.99.B1 |
KCl |
60 mM, the activity increases 1.3-fold, 100 mM KCl result in significant inhibition of activity |
Pyrococcus horikoshii |
|
3.1.99.B1 |
Mg2+ |
addition of 1 mM MgCl2 increases the activity 1.4-fold. 10 mM MgCl2 results in greater than 90% inhibition |
Pyrococcus horikoshii |
|
3.1.99.B1 |
Mn2+ |
addition of 0.1 mM MgCl2 increases the activity 1.5-fold. 10 mM MnCl2 results in greater than 90% inhibition |
Pyrococcus horikoshii |
|
Molecular Weight [Da]
EC Number |
Molecular Weight [Da] |
Molecular Weight Maximum [Da] |
Comment |
Organism |
---|
3.1.99.B1 |
41000 |
- |
gel filtration |
Pyrococcus horikoshii |
Organism
EC Number |
Organism |
UniProt |
Comment |
Textmining |
---|
3.1.99.B1 |
Pyrococcus horikoshii |
O50123 |
- |
- |
Purification (Commentary)
EC Number |
Purification (Comment) |
Organism |
---|
3.1.99.B1 |
- |
Pyrococcus horikoshii |
Storage Stability
EC Number |
Storage Stability |
Organism |
---|
3.1.99.B1 |
4°C or 20°C, stable for over 3 months without loss of activity |
Pyrococcus horikoshii |
Substrates and Products (Substrate)
EC Number |
Substrates |
Comment Substrates |
Organism |
Products |
Comment (Products) |
Rev. |
Reac. |
---|
3.1.99.B1 |
additional information |
the enzyme cleaves replication fork-like substrates and 5' double-strand flap structures using both flap endonuclease and 5'->3'-exonuclease activities |
Pyrococcus horikoshii |
? |
- |
? |
|
Subunits
EC Number |
Subunits |
Comment |
Organism |
---|
3.1.99.B1 |
monomer |
1 * 41000, SDS-PAGE |
Pyrococcus horikoshii |
Temperature Optimum [°C]
EC Number |
Temperature Optimum [°C] |
Temperature Optimum Maximum [°C] |
Comment |
Organism |
---|
3.1.99.B1 |
additional information |
- |
the optimal temperature could not be determined since the activity peak could not be detected |
Pyrococcus horikoshii |
Temperature Stability [°C]
EC Number |
Temperature Stability Minimum [°C] |
Temperature Stability Maximum [°C] |
Comment |
Organism |
---|
3.1.99.B1 |
additional information |
- |
thermostability is dependent on the concentration of NaCl. Addition of 1 M NaCl increases the thermostability |
Pyrococcus horikoshii |
3.1.99.B1 |
95 |
- |
at a final concentration of 0.3 mg/ml, the half-life is 2 h in the presence of 1 M NaCl and 20 min in 50 mM NaCl |
Pyrococcus horikoshii |
3.1.99.B1 |
102 |
- |
Tm-value is 102°C in 0.0085 and 0.0014 mM protein with 1 M NaCl concentrations. Values of Tm are not dependent on the protein concentration |
Pyrococcus horikoshii |
pH Optimum
EC Number |
pH Optimum Minimum |
pH Optimum Maximum |
Comment |
Organism |
---|
3.1.99.B1 |
6 |
7 |
- |
Pyrococcus horikoshii |
pH Range
EC Number |
pH Minimum |
pH Maximum |
Comment |
Organism |
---|
3.1.99.B1 |
6 |
7 |
the activity of phFEN decreases immediately at pH values below 6.0 and over 7.0. The activity can not be detected at pH 5.0 and decreased to 24% at pH 9.4 |
Pyrococcus horikoshii |
pI Value
EC Number |
Organism |
Comment |
pI Value Maximum |
pI Value |
---|
3.1.99.B1 |
Pyrococcus horikoshii |
the pI value of the phFEN is greater than 9.3 |
- |
additional information |