Affinity cross-linked delta-opioid receptor in NG108-15 cells is low molecular weight (25 kDa) and coupled to GTP-binding proteins.

Abstract

The affinity cross-linking of the delta-opioid receptor in neuroblastoma x glioma NG108-15 cells was undertaken using (3-[125I]iodotyrosyl27)human-beta-endorphin ([125I]beta-endorphin) and disuccinimidyl suberate (DSS) or bis(sulfosuccinimidyl) suberate (BS3) in order to estimate molecular size. Following sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis, two radioactive bands were observed. Labeling of a major band of 29 kDa diminished in the presence of unlabeled selective delta-opioid agonist, [D-Pen2,D-Pen5]enkephalin (DPDPE), in a concentration-dependent manner, while labeling of a minor band of 58 kDa was hardly affected. The labeling intensity of the 29 kDa band decreased by addition of guanosine 5'-(3-o-thio)triphosphate (GTP gamma S) or by pretreatment of cells with pertussis toxin. These results, taking the molecular weight of covalently bound beta-endorphin (3.6 kDa) into consideration, suggest that the delta-opioid receptor in NG108-15 cell membrane is a 25 kDa protein which is coupled to pertussis toxin-sensitive guanosine triphosphate-binding proteins (G-proteins).

• Authors: J Harada, H Ueda, Y Iso, M Satoh
• Journal: European journal of pharmacology (Eur J Pharmacol) Vol. 227 Issue 3 Pg. 301-7 (Nov 02 1992) ISSN: 0014-2999 [Print] Netherlands
• PMID: 1335416 (Publication Type: Journal Article)

Chemical References

• Receptors, Opioid
• Succinimides
• Virulence Factors, Bordetella
• beta-Endorphin
• Pertussis Toxin
• GTP-Binding Proteins
• disuccinimidyl suberate

Topics

• Cell Membrane (chemistry)
• Chemical Fractionation
• Electrophoresis, Polyacrylamide Gel
• GTP-Binding Proteins (metabolism)
• Glioma (metabolism)
• Humans
• Molecular Weight
• Neuroblastoma (metabolism)
• Pertussis Toxin
• Receptors, Opioid (chemistry, metabolism)
• Succinimides (pharmacology)
• Tumor Cells, Cultured (metabolism)
• Virulence Factors, Bordetella (pharmacology)
• beta-Endorphin (pharmacology)