Divergent Label-free Cell Phenotypic Pharmacology of Ligands at the Overexpressed beta 2-Adrenergic Receptors

作者：Ferrie Ann M; Sun Haiyan; Zaytseva Natalya; Fang Ye^*

来源：Scientific Reports, 2014, 4(1): 3828.

DOI：10.1038/srep03828

We present subclone sensitive cell phenotypic pharmacology of ligands at the beta(2)-adrenergic receptor (beta(2)-AR) stably expressed in HEK-293 cells. The parental cell line was transfected with green fluorescent protein (GFP)-tagged beta(2)-AR. Four stable subclones were established and used to profile a library of sixty-nine AR ligands. Dynamic mass redistribution (DMR) profiling resulted in a pharmacological activity map suggesting that HEK293 endogenously expresses functional Gi-coupled alpha(2)-AR and G(s)-coupled beta(2)-AR, and the label-free cell phenotypic activity of AR ligands are subclone dependent. Pathway deconvolution revealed that the DMR of epinephrine is originated mostly from the remodeling of actin microfilaments and adhesion complexes, to less extent from the microtubule networks and receptor trafficking, and certain agonists displayed different efficacy towards the cAMP-Epac pathway. We demonstrate that receptor signaling and ligand pharmacology is sensitive to the receptor expression level, and the organization of the receptor and its signaling circuitry.

• 出版日期2014-1-23