Literature summary extracted from
Kravchuk, A.V.; Zhao, L.; Bruzik, K.S.; Tsai, M.D.
Engineering a catalytic metal binding site into a calcium-independent phosphatidylinositol-specific phospholipase C leads to enhanced stereoselectivity (2003), Biochemistry, 42, 2422-2430.
Protein Variants
EC Number |
Protein Variants |
Comment |
Organism |
---|
4.6.1.13 |
D33N/R69D |
PI-PLC double mutant, 50fold activation by 1 mM Ca2+ |
Bacillus sp. (in: Bacteria) |
4.6.1.13 |
R69A |
mutation of the catalytic Arg-69, not activated by Ca2+ |
Bacillus sp. (in: Bacteria) |
4.6.1.13 |
R69C |
mutation of the catalytic Arg-69, not activated by Ca2+ |
Bacillus sp. (in: Bacteria) |
4.6.1.13 |
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) |
4.6.1.13 |
R69E |
mutation of the catalytic Arg-69, inactive mutant, not activated by Ca2+ |
Bacillus sp. (in: Bacteria) |
4.6.1.13 |
R69G |
mutation of the catalytic Arg-69, not activated by Ca2+ |
Bacillus sp. (in: Bacteria) |
4.6.1.13 |
R69N |
mutation of the catalytic Arg-69, not activated by Ca2+ |
Bacillus sp. (in: Bacteria) |
Inhibitors
EC Number |
Inhibitors |
Comment |
Organism |
Structure |
---|
4.6.1.13 |
Cd2+ |
severe inhibition of wild-type PI-PLC |
Bacillus sp. (in: Bacteria) |
|
4.6.1.13 |
Co2+ |
severe inhibition of wild-type PI-PLC |
Bacillus sp. (in: Bacteria) |
|
4.6.1.13 |
Ni2+ |
severe inhibition of wild-type PI-PLC |
Bacillus sp. (in: Bacteria) |
|
4.6.1.13 |
Zn2+ |
severe inhibition of wild-type PI-PLC, complete inhibition of R69D mutant below 1 mM |
Bacillus sp. (in: Bacteria) |
|
Metals/Ions
EC Number |
Metals/Ions |
Comment |
Organism |
Structure |
---|
4.6.1.13 |
Ca2+ |
41fold activation of R69D mutant, slight activation of wild-type PI-PLC |
Bacillus sp. (in: Bacteria) |
|
4.6.1.13 |
Cd2+ |
activates R69D mutant at low concentrations, no activation of wild-type PI-PLC |
Bacillus sp. (in: Bacteria) |
|
4.6.1.13 |
Co2+ |
activates R69D mutant at low concentrations, no activation of wild-type PI-PLC |
Bacillus sp. (in: Bacteria) |
|
4.6.1.13 |
Li+ |
5fold activation of R69D mutant, slight activation of wild-type PI-PLC |
Bacillus sp. (in: Bacteria) |
|
4.6.1.13 |
Mg2+ |
35fold activation of R69D mutant, slight activation of wild-type PI-PLC |
Bacillus sp. (in: Bacteria) |
|
4.6.1.13 |
Mn2+ |
activates R69D mutant at low concentrations, slight activation of wild-type PI-PLC |
Bacillus sp. (in: Bacteria) |
|
4.6.1.13 |
additional information |
wild-type PI-PLC is calcium-independent |
Bacillus sp. (in: Bacteria) |
|
4.6.1.13 |
Sr2+ |
9fold activation of R69D mutant, slight activation of wild-type PI-PLC |
Bacillus sp. (in: Bacteria) |
|
Organism
EC Number |
Organism |
UniProt |
Comment |
Textmining |
---|
4.6.1.13 |
Bacillus sp. (in: Bacteria) |
- |
- |
- |
Purification (Commentary)
EC Number |
Purification (Comment) |
Organism |
---|
4.6.1.13 |
PI-PLC mutants |
Bacillus sp. (in: Bacteria) |
Reaction
EC Number |
Reaction |
Comment |
Organism |
Reaction ID |
---|
4.6.1.13 |
1-phosphatidyl-1D-myo-inositol = 1D-myo-inositol 1,2-cyclic phosphate + 1,2-diacyl-sn-glycerol |
mechanism |
Bacillus sp. (in: Bacteria) |
|
Specific Activity [micromol/min/mg]
EC Number |
Specific Activity Minimum [µmol/min/mg] |
Specific Activity Maximum [µmol/min/mg] |
Comment |
Organism |
---|
4.6.1.13 |
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) |
4.6.1.13 |
3560 |
- |
pH 7.5, 37°C, wild-type PI-PLC, in absence of Ca2+ |
Bacillus sp. (in: Bacteria) |
Substrates and Products (Substrate)
EC Number |
Substrates |
Comment Substrates |
Organism |
Products |
Comment (Products) |
Rev. |
Reac. |
---|
4.6.1.13 |
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]
EC Number |
Temperature Optimum [°C] |
Temperature Optimum Maximum [°C] |
Comment |
Organism |
---|
4.6.1.13 |
37 |
- |
assay at |
Bacillus sp. (in: Bacteria) |
pH Optimum
EC Number |
pH Optimum Minimum |
pH Optimum Maximum |
Comment |
Organism |
---|
4.6.1.13 |
7.5 |
- |
assay at |
Bacillus sp. (in: Bacteria) |