Literature summary for 4.6.1.13 extracted from

Shi, X.; Shao, C.; Zhang, X.; Zambonelli, C.; Redfield, A.G.; Head, J.F.; Seaton, B.A.; Roberts, M.F.
Modulation of Bacillus thuringiensis phosphatidylinositol-specific phospholipase C activity by mutations in the putative dimerization interface (2009), J. Biol. Chem., 284, 15607-15618.

Cloned(Commentary)

Cloned (Comment)	Organism
wild-type and mutants overexpressed in Escherichia coli	Bacillus thuringiensis

Crystallization (Commentary)

Crystallization (Comment)	Organism
mutant Y247S/Y251S in the absence and presence of myo-inositol as well as mutant Y246S/Y247S/Y248S/Y251S, both mutant proteins crystallize as monomers, are very similar to one another, and have no change in the active site region	Bacillus thuringiensis

Protein Variants

Protein Variants	Comment	Organism
additional information	replacing two tyrosines have small effects on enzyme activity. Removal of three or four tyrosine residues weakens binding to phosphatidylcholine surfaces and reduces phosphatidylinositol cleavage by the enzyme as well as phosphatidylcholine activation of inositol 1,2-(cyclic)-phosphate hydrolysis	Bacillus thuringiensis
Y246S/Y247S/Y248S	less active toward phosphatidylinositol solubilized in diheptanoylphosphatidylcholine and when changing the detergent matrix to Triton X-100, as the wild-type	Bacillus thuringiensis
Y246S/Y247S/Y248S/Y251S	less active toward phosphatidylinositol solubilized in diheptanoylphosphatidylcholine and when changing the detergent matrix to Triton X-100, as the wild-type	Bacillus thuringiensis
Y247S/Y251S	exhibits specific activity toward phosphatidylinositol solubilized in diheptanoylphosphatidylcholine comparable to wild-type. Reduced specific activity, when changing the detergent matrix to Triton X-100	Bacillus thuringiensis

Organism

Organism	UniProt	Comment	Textmining
Bacillus thuringiensis	P08954	-	-

Purification (Commentary)

Purification (Comment)	Organism
wild-type and mutants, by gel filtration	Bacillus thuringiensis

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
0.2	-	mutant Y246S/Y247S/Y248S, with 8 mM cIP as substrate	Bacillus thuringiensis
0.4	-	mutant Y246S/Y247S/Y248S/Y251S, with 8 mM cIP as substrate	Bacillus thuringiensis
0.6	-	mutant Y247S/Y251S, with 8 mM cIP as substrate	Bacillus thuringiensis
0.8	-	mutant Y246S/Y247S/Y248S/Y251S, with 8 mM cIP as substrate, in the presence of 5 mM diheptanoylphosphatidylcholine	Bacillus thuringiensis
1.6	-	mutant Y246S/Y247S/Y248S/Y251S, with phosphatidylinositol as substrate, in the presence of 2 mM POPC (for the small unilamellar vesicles)	Bacillus thuringiensis
2.1	-	mutant Y246S/Y247S/Y248S, with phosphatidylinositol as substrate, in the presence of 2 mM POPC (for the small unilamellar vesicles)	Bacillus thuringiensis
2.3	-	wild-type, with 8 mM cIP as substrate	Bacillus thuringiensis
2.9	-	mutant Y246S/Y247S/Y248S, with 8 mM cIP as substrate, in the presence of 5 mM diheptanoylphosphatidylcholine	Bacillus thuringiensis
6.7	-	mutant Y247S/Y251S, with phosphatidylinositol as substrate, in the presence of 2 mM POPC (for the small unilamellar vesicles)	Bacillus thuringiensis
9.5	-	wild-type, with phosphatidylinositol as substrate, in the presence of 2 mM POPC (for the small unilamellar vesicles)	Bacillus thuringiensis
41	-	mutant Y247S/Y251S, with 8 mM cIP as substrate, in the presence of 5 mM diheptanoylphosphatidylcholine	Bacillus thuringiensis
62	-	mutant Y246S/Y247S/Y248S/Y251S, with phosphatidylinositol as substrate, in the presence of 16 mM Triton X-100	Bacillus thuringiensis
73	-	wild-type, with 8 mM cIP as substrate, in the presence of 5 mM diheptanoylphosphatidylcholine	Bacillus thuringiensis
98	-	mutant Y246S/Y247S/Y248S, with phosphatidylinositol as substrate, in the presence of 16 mM Triton X-100	Bacillus thuringiensis
112	-	mutant Y246S/Y247S/Y248S/Y251S, with phosphatidylinositol as substrate, in the presence of 32 mM diheptanoylphosphatidylcholine	Bacillus thuringiensis
239	-	mutant Y247S/Y251S, with phosphatidylinositol as substrate, in the presence of 16 mM Triton X-100	Bacillus thuringiensis
301	-	mutant Y246S/Y247S/Y248S, with phosphatidylinositol as substrate, in the presence of 32 mM diheptanoylphosphatidylcholine	Bacillus thuringiensis
375	-	wild-type, with phosphatidylinositol as substrate, in the presence of 16 mM Triton X-100	Bacillus thuringiensis
560	-	wild-type, with phosphatidylinositol as substrate, in the presence of 32 mM diheptanoylphosphatidylcholine	Bacillus thuringiensis
670	-	mutant Y247S/Y251S, with phosphatidylinositol as substrate, in the presence of 32 mM diheptanoylphosphatidylcholine	Bacillus thuringiensis

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
phosphatidylinositol	-	Bacillus thuringiensis	diacylglycerol + myo-inositol 1,2-cyclic phosphate	-	?

Synonyms

Synonyms	Comment	Organism
phosphatidylinositol-specific phospholipase C	-	Bacillus thuringiensis
PI-PLC	-	Bacillus thuringiensis