Dataset Information

HEK293 femtomolar GPCR signaling

ABSTRACT: G protein-coupled receptors (GPCRs) are the largest class of cell surface signaling proteins; they participate in all physiological processes and are the targets of 30% of marketed drugs. Typically, nanomolar-micromolar concentrations of ligand are used to activate GPCRs in experimental systems. However, by measuring cAMP with increased spatial and temporal resolution, we can now detect GPCR responses to an extraordinarily wide range of ligand concentrations: from attomolar to millimolar. Mathematical modeling shows that the addition of femtomolar concentrations of ligand can activate a significant proportion of cells provided that a cell can be activated by 1-2 binding events. In addition to cAMP, activation of the endogenous b2-adrenoceptor (b2AR) and muscarinic M3R by femtomolar concentrations of ligand in cell lines and human cardiac fibroblasts causes sustained increases in nuclear ERK or cytosolic PKC, respectively. These responses are spatially and temporally distinct from those that occur at higher concentrations of ligand, and result in a unique proteomic profile. This highly sensitive signaling is dependent on the GPCRs forming pre-assembled higher-order signaling complexes at the plasma membrane. Recognizing that GPCRs respond to ultra-low concentrations of neurotransmitters and hormones challenges established paradigms of drug action and provides a new dimension of GPCR activation that is quite distinct from that typically observed.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Kidney Epithelial Cell, Cell Culture

DISEASE(S): Disease Free

SUBMITTER: Srgjan Chivchiristov

LAB HEAD: Michelle Louise Halls

PROVIDER: PXD011033 | Pride | 2018-10-11

REPOSITORIES: Pride

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Preassembled GPCR signaling complexes mediate distinct cellular responses to ultralow ligand concentrations.

Civciristov Srgjan S Ellisdon Andrew M AM Suderman Ryan R Pon Cindy K CK Evans Bronwyn A BA Kleifeld Oded O Charlton Steven J SJ Hlavacek William S WS Canals Meritxell M Halls Michelle L ML

Science signaling 20181009 551

G protein-coupled receptors (GPCRs) are the largest class of cell surface signaling proteins, participate in nearly all physiological processes, and are the targets of 30% of marketed drugs. Typically, nanomolar to micromolar concentrations of ligand are used to activate GPCRs in experimental systems. We detected GPCR responses to a wide range of ligand concentrations, from attomolar to millimolar, by measuring GPCR-stimulated production of cyclic adenosine monophosphate (cAMP) with high spatial ...[more]

PMID: 30301787

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			Action	DRS
	HUMAN.fasta	Fasta
	Legendforbiologicalreplicates.pdf	Pdf
	OK20140117_SC3_pH1.raw	Raw
	OK20140117_SC3_pH11.raw	Raw
	OK20140117_SC3_pH3.raw	Raw