Biomaterial Database

Improved purification and biochemical properties of phosphatidylinositol-specific phospholipase C from Bacillus thuringiensis. 1989

T Kupke, and M Lechner, and G Kaim, and F Götz

Institut für Mikrobielle Genetik, Universität Tübingen, Federal Republic of Germany.

Monophosphatidylinositol inositol phosphohydrolase (phosphatidylinositol-specific phospholipase C. PtdIns-PLC. EC 3.1.4.10) has been purified from a Bacillus thuringiensis culture supernatant and from the cellular fraction of a recombinant Escherichia coli clone containing the PtdIns-PLC gene from B. thuringiensis. The two-step purification procedure involved ion-exchange chromatography on DEAE-Sepharose followed by separation on a Mono-Q/FPLC-column with yields of 32% and 50%, respectively. The molecular mass was determined to be 34 kDa by SDS/PAGE. The isoelectric point of the enzyme was 5.15. The amino-terminal sequences were shown to be identical for the enzymes purified from both organisms. PtdIns-PLC was inhibited by divalent cations using mixed micelles of Triton X-100 and pure phosphatidylinositol. PtdIns-PLC activity was detectable on polyacrylamide gels by activity staining on phosphatidylinostiol-containing agarose.

UI	MeSH Term	Description	Entries
D010727	Phosphoric Diester Hydrolases	A class of enzymes that catalyze the hydrolysis of one of the two ester bonds in a phosphodiester compound. EC 3.1.4.	Phosphodiesterase,Phosphodiesterases,Hydrolases, Phosphoric Diester
D006863	Hydrogen-Ion Concentration	The normality of a solution with respect to HYDROGEN ions; H+. It is related to acidity measurements in most cases by pH	pH,Concentration, Hydrogen-Ion,Concentrations, Hydrogen-Ion,Hydrogen Ion Concentration,Hydrogen-Ion Concentrations
D000595	Amino Acid Sequence	The order of amino acids as they occur in a polypeptide chain. This is referred to as the primary structure of proteins. It is of fundamental importance in determining PROTEIN CONFORMATION.	Protein Structure, Primary,Amino Acid Sequences,Sequence, Amino Acid,Sequences, Amino Acid,Primary Protein Structure,Primary Protein Structures,Protein Structures, Primary,Structure, Primary Protein,Structures, Primary Protein
D000596	Amino Acids	Organic compounds that generally contain an amino (-NH2) and a carboxyl (-COOH) group. Twenty alpha-amino acids are the subunits which are polymerized to form proteins.	Amino Acid,Acid, Amino,Acids, Amino
D001413	Bacillus thuringiensis	A species of gram-positive bacteria which may be pathogenic for certain insects. It is used for the biological control of the Gypsy moth.	Bacilan,Dipel,Thuricide
D043265	Phosphatidylinositol Diacylglycerol-Lyase	A phosphorus-oxygen lyase found primarily in BACTERIA. The enzyme catalyzes the cleavage of a phosphoester linkage in 1-phosphatidyl-1D-myo-inositol to form 1D-myo-inositol 1,2-cyclic phosphate and diacylglycerol. The enzyme was formerly classified as a phosphoric diester hydrolase (EC 3.1.4.10) and is often referred to as a TYPE C PHOSPHOLIPASES. However it is now known that a cyclic phosphate is the final product of this enzyme and that water does not enter into the reaction.	1-Phosphatidylinositol Phosphodiesterase,Monophosphatidylinositol Phosphodiesterase,1 Phosphatidylinositol Phosphodiesterase,Diacylglycerol-Lyase, Phosphatidylinositol,Phosphatidylinositol Diacylglycerol Lyase,Phosphodiesterase, 1-Phosphatidylinositol,Phosphodiesterase, Monophosphatidylinositol
D054801	Phosphoinositide Phospholipase C	A type C phospholipase with specificity towards PHOSPHATIDYLINOSITOLS that contain INOSITOL 1,4,5-TRISPHOSPHATE. Many of the enzymes listed under this classification are involved in intracellular signaling.	1-Phosphatidyl-D-myo-Inositol-4,5-bisphosphate Inositoltrisphosphohydrolase,1-Phosphatidylinositol-4,5-Bisphosphate Phosphodiesterase,PIP2-PLC,PIP2-Phosphoinositidase C,PPI Phosphodiesterase,Phosphatidylinositide-4,5-Bisphosphate Phospholipase C,Phosphatidylinositol-4,5-Biphosphate-Phosphodiesterase,Phosphatidylinositol-Specific Phospholipase C,Phosphoinositide Selective Phospholipase C,Phosphoinositol Specific Phospholipase C,Polyphosphoinositide Phosphodiesterase,Polyphosphoinositide Phospholipase C,PtdIns(4,5)P2 Phosphodiesterase,Tri-phosphoinositide Phosphodiesterase,Triphosphoinositide Phosphodiesterase,1 Phosphatidyl D myo Inositol 4,5 bisphosphate Inositoltrisphosphohydrolase,1 Phosphatidylinositol 4,5 Bisphosphate Phosphodiesterase,Inositoltrisphosphohydrolase, 1-Phosphatidyl-D-myo-Inositol-4,5-bisphosphate,PIP2 Phosphoinositidase C,Phosphatidylinositide 4,5 Bisphosphate Phospholipase C,Phosphatidylinositol 4,5 Biphosphate Phosphodiesterase,Phosphatidylinositol Specific Phospholipase C,Phosphodiesterase, 1-Phosphatidylinositol-4,5-Bisphosphate,Phosphodiesterase, PPI,Phosphodiesterase, Polyphosphoinositide,Phosphodiesterase, Triphosphoinositide,Phospholipase C, Phosphatidylinositide-4,5-Bisphosphate,Phospholipase C, Phosphatidylinositol-Specific,Phospholipase C, Polyphosphoinositide,Tri phosphoinositide Phosphodiesterase