Activating Compound | Comment | Organism | Structure |
---|---|---|---|
KCl | ionic strength, and not the salt identity, is important for PI-PLC activation towards phosphatidylinositol in micelles. Added salt has a synergistic effect with zwitterionic phospholipids, leading to high specific activities for phosphatidylinositol cleavage with only moderate dilution of the anionic substrate in the interface. This kinetic activation correlates with weakening of strong PI-PLC hydrophobic interactions with the interface. PI-PLC cleavage of phosphatidylinositol presented in small unilamellar vesicles is activated by salt | Listeria monocytogenes |
Cloned (Comment) | Organism |
---|---|
inserted into the pTYB11 N-terminal fusion vector between Sap I and EcoR I restriction sites. Recombinant plasmid, IMPACT-Y, transformed into Escherichia coli ER2566 cells for overexpression | Listeria monocytogenes |
Protein Variants | Comment | Organism |
---|---|---|
F237A | most approaches wild-type PI-PLC in its dependence on enzyme concentration | Listeria monocytogenes |
F237W | even at high concentrations, has high specific activity comparable to dilute unaltered recombinant PI-PLC, does not form the aggregates with anionic lipid-rich vesicles that are disrupted by excess detergent and salt, although it is still activated to about the same extent as wild-type by salt | Listeria monocytogenes |
L151A | added KCl (0.15 M) still enhances PI-PLC cleavage of phosphatidylinositol in TX-100 micelles, although KCl effects are much more modest (1.6fold increase) compared to wild-type, F237A or F237W | Listeria monocytogenes |
L235A | added KCl (0.15 M) still enhances PI-PLC cleavage of phosphatidylinositol in TX-100 micelles, although KCl effects are much more modest (1.4fold increase) compared to wild-type, F237A or F237W | Listeria monocytogenes |
W49A | added KCl (0.15 M) still enhances PI-PLC cleavage of phosphatidylinositol in TX-100 micelles, although KCl effects are much more modest (1.8fold increase) compared to wild-type, F237A or F237W | Listeria monocytogenes |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
28000 | - |
1 * 28000, in solution, laser light scattering | Listeria monocytogenes |
28200 | - |
laser light scattering | Listeria monocytogenes |
33000 | - |
sequence analysis | Listeria monocytogenes |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Listeria monocytogenes | - |
- |
- |
Purification (Comment) | Organism |
---|---|
by sonication and centrifugation, on chitin column and by gel filtration, more than 85% pure | Listeria monocytogenes |
Storage Stability | Organism |
---|---|
4°C, 1 month | Listeria monocytogenes |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
dibutyrylphosphatidylinositol | is a poor substrate, necessitating long incubation times (2 to 5 h) if the same enzyme concentration is to be used in the absence and presence of salts and amphiphiles | Listeria monocytogenes | 1D-myo-inositol 1,2-(cyclic)-phosphate + 1,2-dibutyryl-sn-glycerol | - |
? | |
additional information | Phe237 is a key residue in the very tight binding of PI-PLC to membranes containing anionic phospholipids | Listeria monocytogenes | ? | - |
? | |
phosphatidylinositol | - |
Listeria monocytogenes | diacylglycerol + myo-inositol 1,2-cyclic phosphate | - |
? |
Subunits | Comment | Organism |
---|---|---|
monomer | 1 * 28000, in solution, laser light scattering | Listeria monocytogenes |
Synonyms | Comment | Organism |
---|---|---|
phosphatidylinositol-specific phospholipase C | - |
Listeria monocytogenes |
PI-PLC | - |
Listeria monocytogenes |