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In vitro synthesis of 32P-labelled phosphatidylinositol 4,5-bisphosphate and its hydrolysis by smooth muscle membrane-bound phospholipase C

Item Type:Article
Title:In vitro synthesis of 32P-labelled phosphatidylinositol 4,5-bisphosphate and its hydrolysis by smooth muscle membrane-bound phospholipase C
Creators Name:Fuelle, H.J. and Hoeer, D. and Lache, W. and Rosenthal, W. and Schultz, G. and Oberdisse, E.
Abstract:For studies of phospholipase C (PLC) activity in cell-free systems, 32P-labelled phosphatidylinositol 4,5-bisphosphate (PIP2) was prepared enzymatically by phosphorylating phosphatidylinositol 4-phosphate (PIP) in the presence of [gamma-32P]ATP using a PIP kinase partially purified from bovine retinae. PLC activity was determined by incubating membranes of DDT1 MF-2 cells with 32P-PIP2 and measuring remaining non-hydrolyzed substrate as well as accumulation of the hydrolysis product, inositol trisphosphate (IP3). Guanine nucleotides stimulated PIP2 hydrolysis and IP3 release. Additional increase in IP3 accumulation was observed with adrenaline plus guanine nucleotides.
Keywords:Cell Line, Cell-Free System, High Pressure Liquid Chromatography, Epinephrine, Guanosine 5'-O-(3-Thiotriphosphate), Guanosine Triphosphate, Hydrolysis, Smooth Muscle, Phosphatidylinositol 4,5-Diphosphate, Phosphatidylinositols, Thionucleotides, Type C Phospholipases, Animals, Cricetinae
Source:Biochemical and Biophysical Research Communications
ISSN:0006-291X
Publisher:Academic Press (U.S.A.)
Volume:145
Number:2
Page Range:673-679
Date:15 June 1987
Official Publication:https://doi.org/10.1016/0006-291X(87)91017-5
PubMed:View item in PubMed

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