Equivalent regulation of wild type and an epitope-tagged variant of Gs alpha by the IP prostanoid receptor following expression in neuroblastoma x glioma hybrid, NG108-15, cells.
NG108-15 cells were transfected to stably express a haemagglutinin epitope-tagged variant of the long isoform of Gs alpha. Clone BST15 expressed this polypeptide at similar levels to the endogenous long isoform of Gs alpha. Treatment of clone BST15 with the IP prostanoid receptor agonist iloprost resulted in down-regulation of both forms of Gs alpha with both dose-effect curves to iloprost and time courses of loss of the two forms of Gs alpha being indistinguishable. These results demonstrate that the IP prostanoid receptor interacts with and regulates the epitope-tagged variant of Gs alpha in an equivalent manner to the unmodified protein and indicates that the epitope-tagged polypeptide can be used to analyse mechanisms of receptor regulation of cellular G-protein levels.
Mullaney
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I
Division of Biochemistry and Molecular Biology, University of Glasgow, Scotland, UK.
Milligan
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G
D000595:Amino Acid Sequence; D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D015536:Down-Regulation / Q000187:drug effects; D000939:Epitopes / Q000235:genetics; D019204:GTP-Binding Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006388:Hemagglutinins / Q000235:genetics; D006822:Hybrid Cells; D016285:Iloprost / Q000494:pharmacology; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000235:genetics; D011982:Receptors, Prostaglandin / Q000819:agonists / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D000939:Epitopes; D006388:Hemagglutinins; D009842:Oligopeptides; D011982:Receptors, Prostaglandin; D019204:GTP-Binding Proteins; D016285:Iloprost