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Protocol for detecting endogenous GPCR activity in primary cell cultures using ONE-GO biosensors

Remi Janicot1, Mikel Garcia-Marcos2

  • 1Department of Biochemistry & Cell Biology, Chobanian & Avedisian School of Medicine, Boston University, Boston, MA 02118, USA.

STAR Protocols
|October 2, 2024
PubMed
Abstract

Keywords:
Biotechnology and bioengineeringCell BiologyCell cultureCell-based AssaysMolecular BiologySignal Transduction

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G-protein coupled receptors are ligand binding receptors that indirectly affect changes in the cell. The actual receptor is a single polypeptide that transverses the cell membrane seven times creating intracellular and extracellular loops. The extracellular loops create a ligand specific pocket which binds to neurotransmitters or hormones. The intracellular loops holds onto the G-protein.
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G protein-coupled receptor (GPCR) signaling plays a crucial role in cell functioning. GPCR desensitization is an equally essential process. It allows cells to respond to changing environments and regain sensitivity to new stimuli while preventing unnecessary stimulation when no longer needed. Prolonged exposure to stimuli leads to GPCR desensitization. It involves blocking the receptors from binding and activating additional G proteins. This inhibits activation of downstream effectors, thereby...