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Updated: Aug 31, 2026

Measuring G-protein-coupled Receptor Signaling via Radio-labeled GTP Binding
Published on: June 9, 2017
Measurement of G protein-coupled receptor-stimulated phospholipase D activity in intact cells
Abstract:
Mammalian phospholipase D (PLD) activity hydrolyzes phosphatidylcholine (PC) into phosphatidic acid (PA) and free choline. This activity can be stimulated by a wide variety of extracellular agonists, including those for G protein-coupled receptors (GPCRs). This chapter outlines a protocol for the measurement of PLD activity in intact cells following stimulation by an extracellular agonist. The protocol takes advantage of a unique property of mammalian PLDs--the ability to substitute a primary alcohol for water in the hydrolytic reaction. This transphosphatidylation reaction results in the formation of a phosphatidylalcohol, which is a specific and unique marker for PLD activity. This protocol is highly sensitive for the detection of PLD activity following the stimulation of intact cells, being a valuable method for studying the regulation of PLD activity in vivo.
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