Quantifying Gq Signaling Using the IP1 Homogenous Time-Resolved Fluorescence (HTRF) Assay
- 13 October 2024
- book chapter
- Published by Springer Nature
- Vol. 2861, 23-32
Abstract
G protein-coupled receptors (GPCRs) play pivotal roles in cellular signaling and can regulate several cellular functions such as proliferation, secretion, protein expression, and cellular metabolism. Coupling of GPCRs to members of the Gq/11 protein family results in activation of inositol trisphosphate (IP3) and accumulation of calcium intracellularly. This protocol chapter outlines a step-by-step guide for utilizing the inositol phosphate-1 (IP1) accumulation assay, a time-resolved fluorescence resonance energy transfer (TR-FRET) method, to investigate Gq-IP3 signaling. The assay serves as a valuable tool for those conducting pharmacological investigations and compound screening targeting this critical cellular pathway. This protocol chapter covers experimental setup, sample preparation, and data analysis, providing researchers with an in-depth guide to explore the pharmacology of Gq-coupled receptors.Keywords
This publication has 11 references indexed in Scilit:
- IP3 receptor signaling and endothelial barrier functionCellular and Molecular Life Sciences, 2017
- The Gq signalling pathway inhibits brown and beige adipose tissueNature Communications, 2016
- Monitoring Gq-coupled receptor response through inositol phosphate quantification with the IP-One assayExpert Opinion on Drug Discovery, 2011
- Comparison on Functional Assays for Gq-Coupled GPCRs by Measuring Inositol Monophospate-1 and Intracellular Calcium in 1536-Well Plate FormatCurrent Chemical Genomics, 2008
- Mammalian G Proteins and Their Cell Type Specific FunctionsPhysiological Reviews, 2005
- Novel Regulation of Calcium Inhibition of the Inositol 1,4,5-trisphosphate Receptor Calcium-release ChannelJournal of General Physiology, 2003
- Time resolved amplification of cryptate emission: a versatile technology to trace biomolecular interactionsReviews in Molecular Biotechnology, 2002
- Regulation of Phosphoinositide-Specific Phospholipase CAnnual Review of Biochemistry, 2001
- Lipid second messenger regulation: the role of diacylglycerol kinases and their relevance to hypertensionJournal of Human Hypertension, 2001
- Regulation of inositol phospholipid and inositol phosphate metabolism in chemoattractant‐activated human polymorphonuclear leukocytesJournal of Cellular Biochemistry, 1987