Divergent label-free cell phenotypic pharmacology of ligands at the overexpressed β₂-adrenergic receptors.
We present subclone sensitive cell phenotypic pharmacology of ligands at the β2-adrenergic receptor (β2-AR) stably expressed in HEK-293 cells. The parental cell line was transfected with green fluorescent protein (GFP)-tagged β2-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 α2-AR and Gs-coupled β2-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.
1] Biochemical Technologies, Science and Technology Division, Corning Incorporated, Corning, NY 14831, United States of America [2].
Ann M
AM
Ferrie
1] Biochemical Technologies, Science and Technology Division, Corning Incorporated, Corning, NY 14831, United States of America [2] [3].
Haiyan
H
Sun
Biochemical Technologies, Science and Technology Division, Corning Incorporated, Corning, NY 14831, United States of America.
Natalya
N
Zaytseva
Biochemical Technologies, Science and Technology Division, Corning Incorporated, Corning, NY 14831, United States of America.
Ye
Y
Fang
D008841:Actin Cytoskeleton / Q000378:metabolism; D000318:Adrenergic beta-Agonists / Q000494:pharmacology*; D015153:Blotting, Western; D000242:Cyclic AMP / Q000378:metabolism; D004837:Epinephrine / Q000494:pharmacology*; D005434:Flow Cytometry; D049452:Green Fluorescent Proteins / Q000378:metabolism; D020662:Guanine Nucleotide Exchange Factors / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D008024:Ligands; D008870:Microtubules / Q000378:metabolism; D010641:Phenotype; D021381:Protein Transport; D018343:Receptors, Adrenergic, beta-2 / Q000737:chemistry* / Q000378:metabolism*; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects*
D000318:Adrenergic beta-Agonists; D020662:Guanine Nucleotide Exchange Factors; D008024:Ligands; C116122:RAPGEF3 protein, human; D018343:Receptors, Adrenergic, beta-2; D043562:Receptors, G-Protein-Coupled; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D000242:Cyclic AMP; D004837:Epinephrine