[(125)I]Orphanin FQ/nociceptin binding in Raji cells Journal Article
| Authors: | Hom, J. S. H.; Goldberg, I.; Mathis, J.; Pan, Y. X.; Brooks, A. I.; Ryan-Moro, J.; Scheinberg, D. A.; Pasternak, G. W. |
| Article Title: | [(125)I]Orphanin FQ/nociceptin binding in Raji cells |
| Abstract: | Western blots using an antibody which recognizes the orphanin FQ/ nociceptin (OFQ/N) receptor reveals a band at approximately 69 kD in several cell lines, including the Raji human B cell lymphoma cell line. RT-PCR confirms the presence of this receptor in the Raji cells. Binding studies revealed a high affinity [125I][Tyr14]OFQ/N site in the Raji cells. The affinity of [125I] [Tyr14]OFQ/N in the Raji cells (K(D) 68.4 pM) was similar to that in the transfected receptor (K(D) 36.7 pM). Its selectivity profile also was quite similar. OFQ/N competed binding quite potently (K(i) 65 pM), as did [Tyr14]OFQ/N (K(i) 33 pM). Traditional opioids displayed no appreciable affinity for the binding at any concentration examined, with the exception of naloxone benzoylhydrazone, which had only a very modest affinity. The receptors in the Raji cells were functionally active. OFQ/N inhibited forskolin-stimulated cyclase by 72% with an IC50 value of approximately 1 nM. |
| Keywords: | human cell; binding affinity; metabolism; reverse transcription polymerase chain reaction; enzyme inhibition; opiate; protein binding; ph; tumor cells, cultured; time; time factors; monoclonal antibody; b cell lymphoma; blotting, western; lymphoma, b-cell; antibodies, monoclonal; iodine 125; radioactive iodine; cell culture; iodine radioisotopes; western blotting; immunoblotting; binding site; binding sites; immunocompetent cell; hydrogen-ion concentration; opiate receptor; enzyme; opioid peptides; naloxone benzoylhydrazone; raji cell; nociceptin; opiate peptide; forskolin; radioassay; radioligand assay; immune cells; receptors, opioid; humans; human; priority journal; article; nociceptin receptor; orl1; orphanin fq; kor-3 |
| Journal Title: | Synapse |
| Volume: | 34 |
| Issue: | 3 |
| ISSN: | 0887-4476 |
| Publisher: | John Wiley & Sons |
| Date Published: | 1999-12-01 |
| Start Page: | 187 |
| End Page: | 191 |
| Language: | English |
| DOI: | 10.1002/(sici)1098-2396(19991201)34:3<187::aid-syn3>3.0.co;2-a |
| PUBMED: | 10523756 |
| PROVIDER: | scopus |
| DOI/URL: | |
| Notes: | Article -- Export Date: 16 August 2016 -- Source: Scopus |
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