Application | Comment | Organism |
---|---|---|
additional information | Gly56 is required in stabilizing the propeptide fold. Stabilization of this fold leads to strong binding of the propeptide to subtilisin, high resistance of the propeptide to proteolytic degradation, and slow maturation of Pro-subtilisin | Thermococcus kodakarensis |
Cloned (Comment) | Organism |
---|---|
pET25b derivatives for overproduction of Pro-G56W, Pro-G56E, G56W-propeptide, and G56E-propeptide. G56W-propeptide overproduced in soluble form, while G56E-propeptide overproduced in insoluble form. S324A-subtilisin overproduced in inclusion bodies | Thermococcus kodakarensis |
Crystallization (Comment) | Organism |
---|---|
by the sitting drop vapor diffusion method. Crystal structures of propeptides in complex with S324A-subtilisin. Conformation of the propeptide is altered by the mutation, such that nonglycine residues at position 56 assume a right-handed conformation and hydrophobic interactions at the core region decrease | Thermococcus kodakarensis |
Protein Variants | Comment | Organism |
---|---|---|
G56E | Pro-G56E gives a halo at temperatures equal or higher than 60 °C | Thermococcus kodakarensis |
G56F | Pro-G56F gives a halo only at temperatures equal or higher than 70 °C | Thermococcus kodakarensis |
G56S | Pro-G56S gives a halo from 40-80°C | Thermococcus kodakarensis |
G56W | Pro-G56W gives a halo only at 80 °C like the wild-type | Thermococcus kodakarensis |
S324A | active-site mutant of Pro-subtilisin | Thermococcus kodakarensis |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
propeptide | inhibition in a concentration-dependent manner. Wild-type propeptide is more potent than G56W-, G56S- and G56E-propeptide | Thermococcus kodakarensis |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Ca2+ | - |
Thermococcus kodakarensis |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Thermococcus kodakarensis | - |
- |
- |
Purification (Comment) | Organism |
---|---|
G56W-propeptide and G56E-propeptide, S324A-subtilisin purified in denatured form | Thermococcus kodakarensis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
casein + H2O | - |
Thermococcus kodakarensis | ? | - |
? | |
N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide + H2O | - |
Thermococcus kodakarensis | N-succinyl-Ala-Ala-Pro-Phe + p-nitroaniline | - |
? |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
40 | 80 | - |
Thermococcus kodakarensis |