Biomaterial Database

Purification of phospholipase C beta and phospholipase C epsilon from Sf9 cells. 2004

Mousumi Ghosh, and Huan Wang, and Grant G Kelley, and Alan V Smrcka

Department of Pharmacology and Physiology, University of Rochester School of Medicine, NY, USA.

Phosphatidylinositol-specific phospholipase C (PLC) enzymes catalyze hydrolysis of phosphatidylinositol 4,5-bisphosphate (PIP2) generating diacylglycerol (DAG) and inositol 1,4,5-triphosphate (IP(3)). The PLC beta isoforms of PLCs are activated by G proteins after hormone or neurotransmitter stimulation of G protein-coupled receptors (GPCR). PLC epsilon is a recently identified PLC isoform that is activated by Ras and G beta gamma subunit although the physiological role of this enzyme is not well understood. Methods for purification of PLC beta and PLC epsilon from Sf9 cells are described. In the case of hexahistidine (6-His)-tagged PLC beta the purification involves two steps, affinity chromatography with Ni-NTA agarose followed by heparin Sepharose chromatography. 6-His-tagged PLC epsilon can be purified in a single step with nickel nitrilotriacetic acid-agarose (Ni-NTA) affinity chromatography.

UI	MeSH Term	Description	Entries
D007313	Insecta	Members of the phylum ARTHROPODA composed or organisms characterized by division into three parts: head, thorax, and abdomen. They are the dominant group of animals on earth with several hundred thousand different kinds. Three orders, HEMIPTERA; DIPTERA; and SIPHONAPTERA; are of medical interest in that they cause disease in humans and animals. (From Borror et al., An Introduction to the Study of Insects, 4th ed, p1).	Insects,Insect
D007527	Isoenzymes	Structurally related forms of an enzyme. Each isoenzyme has the same mechanism and classification, but differs in its chemical, physical, or immunological characteristics.	Alloenzyme,Allozyme,Isoenzyme,Isozyme,Isozymes,Alloenzymes,Allozymes
D010738	Type C Phospholipases	A subclass of phospholipases that hydrolyze the phosphoester bond found in the third position of GLYCEROPHOSPHOLIPIDS. Although the singular term phospholipase C specifically refers to an enzyme that catalyzes the hydrolysis of PHOSPHATIDYLCHOLINE (EC 3.1.4.3), it is commonly used in the literature to refer to broad variety of enzymes that specifically catalyze the hydrolysis of PHOSPHATIDYLINOSITOLS.	Lecithinase C,Phospholipase C,Phospholipases, Type C,Phospholipases C
D011994	Recombinant Proteins	Proteins prepared by recombinant DNA technology.	Biosynthetic Protein,Biosynthetic Proteins,DNA Recombinant Proteins,Recombinant Protein,Proteins, Biosynthetic,Proteins, Recombinant DNA,DNA Proteins, Recombinant,Protein, Biosynthetic,Protein, Recombinant,Proteins, DNA Recombinant,Proteins, Recombinant,Recombinant DNA Proteins,Recombinant Proteins, DNA
D002460	Cell Line	Established cell cultures that have the potential to propagate indefinitely.	Cell Lines,Line, Cell,Lines, Cell
D002846	Chromatography, Affinity	A chromatographic technique that utilizes the ability of biological molecules, often ANTIBODIES, to bind to certain ligands specifically and reversibly. It is used in protein biochemistry. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed)	Chromatography, Bioaffinity,Immunochromatography,Affinity Chromatography,Bioaffinity Chromatography
D000818	Animals	Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA.	Animal,Metazoa,Animalia
D015398	Signal Transduction	The intracellular transfer of information (biological activation/inhibition) through a signal pathway. In each signal transduction system, an activation/inhibition signal from a biologically active molecule (hormone, neurotransmitter) is mediated via the coupling of a receptor/enzyme to a second messenger system or to an ion channel. Signal transduction plays an important role in activating cellular functions, cell differentiation, and cell proliferation. Examples of signal transduction systems are the GAMMA-AMINOBUTYRIC ACID-postsynaptic receptor-calcium ion channel system, the receptor-mediated T-cell activation pathway, and the receptor-mediated activation of phospholipases. Those coupled to membrane depolarization or intracellular release of calcium include the receptor-mediated activation of cytotoxic functions in granulocytes and the synaptic potentiation of protein kinase activation. Some signal transduction pathways may be part of larger signal transduction pathways; for example, protein kinase activation is part of the platelet activation signal pathway.	Cell Signaling,Receptor-Mediated Signal Transduction,Signal Pathways,Receptor Mediated Signal Transduction,Signal Transduction Pathways,Signal Transduction Systems,Pathway, Signal,Pathway, Signal Transduction,Pathways, Signal,Pathways, Signal Transduction,Receptor-Mediated Signal Transductions,Signal Pathway,Signal Transduction Pathway,Signal Transduction System,Signal Transduction, Receptor-Mediated,Signal Transductions,Signal Transductions, Receptor-Mediated,System, Signal Transduction,Systems, Signal Transduction,Transduction, Signal,Transductions, Signal
D054799	Phospholipase C beta	A phosphoinositide phospholipase C subtype that is primarily regulated by its association with HETEROTRIMERIC G-PROTEINS. It is structurally related to PHOSPHOLIPASE C DELTA with the addition of C-terminal extension of 400 residues.	PLC beta,PLC beta1,PLC beta2,PLC beta3,PLC beta4,Phospholipase C beta1,Phospholipase C beta2,Phospholipase C beta3,Phospholipase C beta4,Phospholipase C-beta,Phospholipase C-beta1,Phospholipase C-beta2,Phospholipase C-beta3,Phospholipase C-beta4
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