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Updated: Jul 10, 2026

A Pipeline to Investigate the Structures and Signaling Pathways of Sphingosine 1-Phosphate Receptors
Published on: June 8, 2022
Analysis and pharmacological targeting of phospholipase C beta interactions with G proteins
David M Lehmann1, Chujun Yuan, Alan V Smrcka
1Department of Pharmacology and Physiology, University of Rochester School of Medicine and Dentistry, Rochester, New York, USA.
Abstract:
Phosphatidylinositol-specific phospholipase C enzymes (PLC) catalyze hydrolysis of phosphatidylinositol 4,5-bisphosphate generating the second messengers diacylglycerol and inositol 1,4,5-triphosphate. Mammalian phosphoinositide-specific phospholipase C beta (PLCbeta) activity is regulated by the alpha(q) family of G-protein alpha subunits and by Gbetagamma subunits. Regulation of PLCbeta enzymatic activity can be assayed by reconstituting purified G-protein subunits with purified PLCbeta in the presence of phospholipid vesicles containing the substrate phosphatidylinositol 4,5-bisphosphate. This chapter describes methods for expression and purification of PLCbeta and Gbetagamma from insect cells, assay of G-protein-dependent regulation of PLC activity, and assessment of G-protein-PLC direct binding interactions. This combination of functional and direct binding analysis provides a powerful approach to characterizing PLC and G-protein interfaces, identifying inhibitors of this interaction, and potentially uncovering new modes of PLC regulation.
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