biostudies-literature00490Kumari PDBT/Wellcome Trust India AllianceWellcome Trust141-153https://www.ebi.ac.uk/biostudies/studies/S-EPMC7115865biostudies-literatureUnknown149Agonist stimulation of G-protein-coupled receptors (GPCRs) typically results in phosphorylation and activation of ERK (Extracellular-signal Regulated Kinase) which is a member of MAP kinase (Mitogen-Activated Protein kinase) family. Detection of phosphorylated ERK1/2 MAP kinase has been widely used as readout of GPCR signaling in heterologous cells, primary cells, tissues and even in animal studies. ERK1/2 phosphorylation downstream of GPCRs is now well established to arise from the activation of both, the heterotrimeric G-proteins and β-arrestins (βarrs) with distinct spatio-temporal components. Here, we present a step-by-step protocol for measuring agonist-induced ERK1/2 MAP kinase activation downstream of GPCRs using standard Western blotting assay. Note: ERK1/2 is also referred to as p44/42 MAP kinase. ERK1 and ERK2 are same as Mitogen-Activated Protein Kinase 3 (MAP3) and Mitogen-Activated Protein Kinase 1 (MAP1), respectively.Methods in cell biologyMeasuring agonist-induced ERK MAP kinase phosphorylation for G-protein-coupled receptors.PMC7115865IA/I/14/1/501285Shukla AKDwivedi HKumari PBaidya M49falseMeasuring agonist-induced ERK MAP kinase phosphorylation for G-protein-coupled receptors.Agonist stimulation of G-protein-coupled receptors (GPCRs) typically results in phosphorylation and activation of ERK (Extracellular-signal Regulated Kinase) which is a member of MAP kinase (Mitogen-Activated Protein kinase) family. Detection of phosphorylated ERK1/2 MAP kinase has been widely used as readout of GPCR signaling in heterologous cells, primary cells, tissues and even in animal studies. ERK1/2 phosphorylation downstream of GPCRs is now well established to arise from the activation of both, the heterotrimeric G-proteins and β-arrestins (βarrs) with distinct spatio-temporal components. Here, we present a step-by-step protocol for measuring agonist-induced ERK1/2 MAP kinase activation downstream of GPCRs using standard Western blotting assay. Note: ERK1/2 is also referred to as p44/42 MAP kinase. ERK1 and ERK2 are same as Mitogen-Activated Protein Kinase 3 (MAP3) and Mitogen-Activated Protein Kinase 1 (MAP1), respectively.2019-01-01T00:00:00Z20192024-11-19T23:57:00.986Z2020-08-28T07:00:55ZS-EPMC71158653061681610.1016/bs.mcb.2018.09.015