Crystallization (Comment) | Organism |
---|---|
the 12-subunit PhTET3 complex is crystallized and its 3D structure is solved at a resolution of 1.9 A by molecular replacement | Pyrococcus horikoshii |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
amastatin | 0.002 mM, 77% inhibition | Pyrococcus horikoshii | |
bestatin | 0.5 mM, 34% inhibition | Pyrococcus horikoshii | |
Ca2+ | 1 mM,% inhibition | Pyrococcus horikoshii | |
Cd2+ | 1 mM,% inhibition | Pyrococcus horikoshii | |
CHAPS buffer | - |
Pyrococcus horikoshii | |
chymostatin | 1 mM, 22% inhibition | Pyrococcus horikoshii | |
Cu2+ | 1 mM,% inhibition | Pyrococcus horikoshii | |
EDTA | 0.02 mM, 85% inhibition | Pyrococcus horikoshii | |
Mg2+ | 1 mM,% inhibition | Pyrococcus horikoshii | |
Mn2+ | 1 mM,% inhibition | Pyrococcus horikoshii | |
additional information | the enzyme is insensitive to the papain and trypsin inhibitor antipain, to the cysteine proteases inhibitor E-64, to the serine and cysteine proteases inhibitor leupeptin, to the aspartate proteases inhibitor pepstatin, and to the metalloendopeptidases inhibitor phosphoramidon. Puromycin, an inhibitor of some exopeptidases, has practically no effect on the activity of PhTET3 | Pyrococcus horikoshii | |
pefabloc | 5 mM, 20% inhibition | Pyrococcus horikoshii | |
Zn2+ | 1 mM,% inhibition | Pyrococcus horikoshii |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Co2+ | 1 mM, 2.4fold activation | Pyrococcus horikoshii |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
40000 | - |
12 * 40000, SDS-PAGE, the active sites of the PhTET complexes are located inside the particles, in the catalytic chambers that are enclosed by the tetrahedron apices. The walls of these chambers are positively charged, and each one contains three active sites arranged in a circular way. Adjacent to the active sites, the specificity pockets appear as small cavities. The geometry of these pockets is different in each PhTET complex, as well as their location in the particles relative to the active sites. In PhTET3, the specificity pocket is formed by the side-chains of D262, T240, T295 and T297, which leads to a negative charge | Pyrococcus horikoshii |
500000 | - |
dodecameric complex, gel filtration | Pyrococcus horikoshii |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pyrococcus horikoshii | O59485 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-alanyl-L-alanine 4-nitroanilide + H2O | L-alanine 4-nitroanilide accumulates very fast in the reaction mixture, and the L-alanine 4-nitroanilide/4-nitroaniline ratio diminishes from 123 at 5 min to 69 at 25 min, therefore indicating that the enzyme is mostly cleaving the substrate to L-alanine 4-nitroanilide, and then also this last product to give 4-nitroaniline | Pyrococcus horikoshii | L-alanine + L-alanine 4-nitroanilide | - |
? | |
L-alanyl-L-alanyl-L-alanine 4-nitroanilide + H2O | L-alanyl-L-alanine 4-nitroanilide is produced, but also consumed by the enzyme while 4-nitroaniline is generated after a lag phase. At 26 min the ratios L-alanyl-L-alanine 4-nitroaniline/4-nitroaniline and L-alanyl 4-nitroanilide/4-nitroaniline are 8 and 33 respectively | Pyrococcus horikoshii | L-alanine + L-alanyl-L-alanine 4-nitroanilide | - |
? | |
L-arginine 4-nitroanilide + H2O | 66.6% of the activity compared to L-lysine 4-nitroanilide | Pyrococcus horikoshii | L-arginine + 4-nitroaniline | - |
? | |
L-glutamate 4-nitroanilide + H2O | 20.1% of the activity compared to L-lysine 4-nitroanilide | Pyrococcus horikoshii | L-glutamate + 4-nitroaniline | - |
? | |
L-leucine 4-nitroanilide + H2O | 11.4% of the activity compared to L-lysine 4-nitroanilide | Pyrococcus horikoshii | L-leucine + 4-nitroaniline | - |
? | |
L-lysine 4-nitroanilide + H2O | - |
Pyrococcus horikoshii | L-lysine + 4-nitroaniline | - |
? | |
additional information | poor activity is detected against L-ornithinyl- and L-glutaminyl-7-amido-4-methylcoumarin, and L-aspartyl-, L-methionyl- and L-alanyl 4-nitroanilide, while the enzyme shows very poor or no activity against the other aminoacyl-4-nitroanilides or aminoacyl-7-amido-4-methylcoumarines assayed. No activity can be measured against any of the N-terminal blocked derivatives | Pyrococcus horikoshii | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
dodecamer | 12 * 40000, SDS-PAGE, the active sites of the PhTET complexes are located inside the particles, in the catalytic chambers that are enclosed by the tetrahedron apices. The walls of these chambers are positively charged, and each one contains three active sites arranged in a circular way. Adjacent to the active sites, the specificity pockets appear as small cavities. The geometry of these pockets is different in each PhTET complex, as well as their location in the particles relative to the active sites. In PhTET3, the specificity pocket is formed by the side-chains of D262, T240, T295 and T297, which leads to a negative charge | Pyrococcus horikoshii |
Synonyms | Comment | Organism |
---|---|---|
PH1821 | - |
Pyrococcus horikoshii |
PhTET3 | - |
Pyrococcus horikoshii |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
80 | - |
half-life: 17.83 min | Pyrococcus horikoshii |
90 | - |
half-life: 3.44 min | Pyrococcus horikoshii |
95 | - |
half-life: 55.1 min | Pyrococcus horikoshii |
100 | - |
half-life: 27 min | Pyrococcus horikoshii |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.4 | - |
- |
Pyrococcus horikoshii |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
6 | 8.5 | pH 6: about 50% of maximal activity, pH 8.0: about 90% of maximal activity | Pyrococcus horikoshii |