Literature summary extracted from
Feng, J.; Bradley, W.D.; Roberts, M.F.
Optimizing the interfacial binding and activity of a bacterial phosphatidylinositol-specific phospholipase C (2003), J. Biol. Chem., 278, 24651-24657.
Activating Compound
EC Number |
Activating Compound |
Comment |
Organism |
Structure |
---|
4.6.1.13 |
diheptanoyl phosphatidylcholine |
activates |
Bacillus thuringiensis |
|
4.6.1.13 |
Isopropanol |
water-miscible, 30%, activates |
Bacillus thuringiensis |
|
4.6.1.13 |
phosphatidylcholine |
PI-PLC is activated by nonsubstrate interfaces such as phosphatidylcholine micelles or bilayers, activation corresponds with partial insertion into the interface of Trp-47 and Trp-242 in the rim of the alphabeta-barrel |
Bacillus thuringiensis |
|
Cloned(Commentary)
EC Number |
Cloned (Comment) |
Organism |
---|
4.6.1.13 |
overexpression in Escherichia coli BL21 |
Bacillus thuringiensis |
Protein Variants
EC Number |
Protein Variants |
Comment |
Organism |
---|
4.6.1.13 |
G238W |
study of the kinetic activation by diheptanoyl phosphatidylcholine and water-miscible isopropanol |
Bacillus thuringiensis |
4.6.1.13 |
G238W/W242A |
double mutant with enhanced activation and affinity for phosphatidylcholine interfaces above that of wild-type PI-PLC |
Bacillus thuringiensis |
4.6.1.13 |
G48W/W47A |
double mutant, study of the kinetic activation by diheptanoyl phosphatidylcholine and water-miscible isopropanol |
Bacillus thuringiensis |
4.6.1.13 |
I43W/W47A |
double mutant with recovered kinetic interfacial activation, lower specific activity than wild-type PI-PLC |
Bacillus thuringiensis |
4.6.1.13 |
M49W/W47A |
double mutant, study of the kinetic activation by diheptanoyl phosphatidylcholine and water-miscible isopropanol |
Bacillus thuringiensis |
4.6.1.13 |
additional information |
tryptophan rescue mutagenesis, reinsertion of a Trp at a different place in helix B in the W47A mutant or in the loop of the W242A mutant |
Bacillus thuringiensis |
4.6.1.13 |
N243W/W242A |
double mutant, study of the kinetic activation by diheptanoyl phosphatidylcholine and water-miscible isopropanol |
Bacillus thuringiensis |
4.6.1.13 |
Q45W/W47A |
double mutant, study of the kinetic activation by diheptanoyl phosphatidylcholine and water-miscible isopropanol |
Bacillus thuringiensis |
4.6.1.13 |
S236W/W242A |
double mutant, study of the kinetic activation by diheptanoyl phosphatidylcholine and water-miscible isopropanol |
Bacillus thuringiensis |
4.6.1.13 |
W242A |
mutant with much weaker binding to interfaces and lower kinetic interfacial activation |
Bacillus thuringiensis |
4.6.1.13 |
W47A |
mutant with much weaker binding to interfaces and lower kinetic interfacial activation |
Bacillus thuringiensis |
Organic Solvent Stability
EC Number |
Organic Solvent |
Comment |
Organism |
---|
4.6.1.13 |
isopropanol |
water-miscible, 30%, activates |
Bacillus thuringiensis |
Organism
EC Number |
Organism |
UniProt |
Comment |
Textmining |
---|
4.6.1.13 |
Bacillus thuringiensis |
- |
- |
- |
Purification (Commentary)
EC Number |
Purification (Comment) |
Organism |
---|
4.6.1.13 |
recombinant PI-PLC |
Bacillus thuringiensis |
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 |
- |
- |
Bacillus thuringiensis |
Substrates and Products (Substrate)
EC Number |
Substrates |
Comment Substrates |
Organism |
Products |
Comment (Products) |
Rev. |
Reac. |
---|
4.6.1.13 |
1-phosphatidyl-1D-myo-inositol |
the active site is located at the C-terminal side |
Bacillus thuringiensis |
1D-myo-inositol 1,2-cyclic phosphate + diacylglycerol |
- |
? |
|
Temperature Stability [°C]
EC Number |
Temperature Stability Minimum [°C] |
Temperature Stability Maximum [°C] |
Comment |
Organism |
---|
4.6.1.13 |
54.4 |
- |
Tm-value, wild-type PI-PLC |
Bacillus thuringiensis |
pH Optimum
EC Number |
pH Optimum Minimum |
pH Optimum Maximum |
Comment |
Organism |
---|
4.6.1.13 |
7 |
- |
assay at |
Bacillus thuringiensis |