Protein Variants | Comment | Organism |
---|---|---|
D33N/R69D | PI-PLC double mutant, 50fold activation by 1 mM Ca2+ | Bacillus sp. (in: Bacteria) |
R69A | mutation of the catalytic Arg-69, not activated by Ca2+ | Bacillus sp. (in: Bacteria) |
R69C | mutation of the catalytic Arg-69, not activated by Ca2+ | Bacillus sp. (in: Bacteria) |
R69D | reduced activity compared with wild-type enzyme, mutant is activated by Ca2+, mutation engineers a catalytic metal binding site into the calcium-independent PI-PLC leading to enhanced stereoselectivity | Bacillus sp. (in: Bacteria) |
R69E | mutation of the catalytic Arg-69, inactive mutant, not activated by Ca2+ | Bacillus sp. (in: Bacteria) |
R69G | mutation of the catalytic Arg-69, not activated by Ca2+ | Bacillus sp. (in: Bacteria) |
R69N | mutation of the catalytic Arg-69, not activated by Ca2+ | Bacillus sp. (in: Bacteria) |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Cd2+ | severe inhibition of wild-type PI-PLC | Bacillus sp. (in: Bacteria) | |
Co2+ | severe inhibition of wild-type PI-PLC | Bacillus sp. (in: Bacteria) | |
Ni2+ | severe inhibition of wild-type PI-PLC | Bacillus sp. (in: Bacteria) | |
Zn2+ | severe inhibition of wild-type PI-PLC, complete inhibition of R69D mutant below 1 mM | Bacillus sp. (in: Bacteria) |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Ca2+ | 41fold activation of R69D mutant, slight activation of wild-type PI-PLC | Bacillus sp. (in: Bacteria) | |
Cd2+ | activates R69D mutant at low concentrations, no activation of wild-type PI-PLC | Bacillus sp. (in: Bacteria) | |
Co2+ | activates R69D mutant at low concentrations, no activation of wild-type PI-PLC | Bacillus sp. (in: Bacteria) | |
Li+ | 5fold activation of R69D mutant, slight activation of wild-type PI-PLC | Bacillus sp. (in: Bacteria) | |
Mg2+ | 35fold activation of R69D mutant, slight activation of wild-type PI-PLC | Bacillus sp. (in: Bacteria) | |
Mn2+ | activates R69D mutant at low concentrations, slight activation of wild-type PI-PLC | Bacillus sp. (in: Bacteria) | |
additional information | wild-type PI-PLC is calcium-independent | Bacillus sp. (in: Bacteria) | |
Sr2+ | 9fold activation of R69D mutant, slight activation of wild-type PI-PLC | Bacillus sp. (in: Bacteria) |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Bacillus sp. (in: Bacteria) | - |
- |
- |
Purification (Comment) | Organism |
---|---|
PI-PLC mutants | Bacillus sp. (in: Bacteria) |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
1-phosphatidyl-1D-myo-inositol = 1D-myo-inositol 1,2-cyclic phosphate + 1,2-diacyl-sn-glycerol | mechanism | Bacillus sp. (in: Bacteria) |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
values for wild-type and several mutant PI-PLCs in presence of 0.1 mM or 1 mM Ca2+ and in absence of Ca2+ | Bacillus sp. (in: Bacteria) |
3560 | - |
pH 7.5, 37°C, wild-type PI-PLC, in absence of Ca2+ | Bacillus sp. (in: Bacteria) |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
1-phosphatidyl-1D-myo-inositol | PLC accepts only nonphosphorylated phosphatidylinositol substrates and produces cyclic inositol phosphate as final product, which is hydrolyzed at a 1000fold lower rate, catalytic mechanism, uses a guanidinium group of Arg-69 during catalysis | Bacillus sp. (in: Bacteria) | 1D-myo-inositol 1,2-cyclic phosphate + diacylglycerol | - |
? |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Bacillus sp. (in: Bacteria) |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Bacillus sp. (in: Bacteria) |