Presentation: 3B04

Several key parameters are used to measure GPCR screening, including Ca²⁺, cAMP and IP3. However, all of these assays have limitations. We have developed a new HTS assay platform for GPCR discovery, which employs detection of phosphorylation of endogenous MAP kinase (ERK 1/2). The ERK 1/2 assay, termed 'SureFire', measures phosphorylation of endogenous cellular ERK 1/2 by MEK in high density plates, and can be used to measure activation of ERK 1/2 in cells carrying cloned or endogenous receptors. The assay utilizes AlphaScreen (PerkinElmer) and is fully homogeneous and robotics compatible. We show that ERK 1/2 is consistently activated in cells stimulated by G_q-coupled GPCRs, providing a new HTS platform for screening of GPCR drug candidates. Furthermore, cellular ERK 1/2 can be used to determine activation of non G_q-coupled pathways including G_i and G_s, providing a single platform for detection of several distinct GPCR signaling pathways.

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