Evidence for opioid receptor-mediated activation of the G-proteins, Go and Gi2, in membranes of neuroblastoma x glioma (NG108-15) hybrid cells.
In membranes of neuroblastoma x glioma (NG108-15) hybrid cells, the photoreactive GTP analog, [alpha-32P] GTP azidoanilide, was incorporated into 39-41-kDa proteins comigrating in urea-containing sodium dodecyl sulfate-polyacrylamide gels with immunologically identified G-protein alpha-subunits, i.e. a 39-kDa Go alpha-subunit, a 40-kDa Gi2 alpha-subunit, and a 41-kDa Gi alpha-subunit of an unknown subtype. The synthetic opioid, D-Ala2,D-Leu5-enkephalin (DADLE), stimulated photolabeling of the 39-41-kDa proteins. In the presence of GDP, which increased the ratio of agonist-stimulated to basal photolabeling, DADLE at a maximally effective concentration stimulated photolabeling of the 39- and the 40-kDa protein 2-3-fold. Somatostatin, adrenaline, and bradykinin were less potent than DADLE and, to varying degrees, stimulated photolabeling of the 40-kDa protein more than that of the 39-kDa protein. Prostaglandin E1 was inactive. The present data represent direct evidence for an activation of endogenous Go and Gi2 via opioid receptors and other receptors in the native membrane milieu.
Offermanns
S
S
Institut für Pharmakologie, Freie Universität Berlin, Federal Republic of Germany.
Schultz
G
G
Rosenthal
W
W
D000527:Alprostadil / Q000494:pharmacology; D001920:Bradykinin / Q000494:pharmacology; D002352:Carrier Proteins / Q000187:drug effects / Q000378:metabolism*; D015815:Cell Adhesion Molecules; D004591:Electrophoresis, Polyacrylamide Gel; D016308:Enkephalin, Leucine-2-Alanine / Q000494:pharmacology; D004837:Epinephrine / Q000494:pharmacology; D019204:GTP-Binding Proteins / Q000378:metabolism*; D005910:Glioma / Q000378:metabolism*; D006822:Hybrid Cells; D008562:Membrane Glycoproteins / Q000187:drug effects / Q000378:metabolism*; D009270:Naloxone / Q000494:pharmacology; D009447:Neuroblastoma / Q000378:metabolism*; D013004:Somatostatin / Q000494:pharmacology; D014407:Tumor Cells, Cultured / Q000378:metabolism
D002352:Carrier Proteins; D015815:Cell Adhesion Molecules; D008562:Membrane Glycoproteins; D009270:Naloxone; D013004:Somatostatin; D016308:Enkephalin, Leucine-2-Alanine; D019204:GTP-Binding Proteins; D000527:Alprostadil; D001920:Bradykinin; D004837:Epinephrine