Immunoglobulin G1 Fc fragment-tagged human opioid receptor-like receptor retains the ability to inhibit cAMP accumulation.
The human opioid receptor-like (ORL(1)) receptor was tagged with the immunoglobulin G1 Fc fragment at the carboxy-terminus and expressed in human embryonic kidney 293 cells. Expression of the ORL(1)-Fc receptor was confirmed by immunohistochemical staining. The fusion protein was enriched by affinity chromatography and then verified by immunodetection. The function of the ORL(1)-Fc receptor was determined by examining nociceptin/OFQ-induced inhibition of cAMP accumulation. The ORL(1)-Fc receptor inhibited the forskolin-stimulated cAMP accumulation. The inhibitory response was selectively induced by nociceptin/OFQ in a dose-dependent and pertussis toxin-sensitive manner. Our results indicate that the carboxy-terminal Fc-tagged ORL(1) receptor retained the ability to interact with G(i) proteins to inhibit adenylyl cyclase.
Yung
L Y
LY
Department of Biology and the Biotechnology Research Institute, Hong Kong University of Science and Technology, Kowloon, Hong Kong, China.
Tsim
K W
KW
Pei
G
G
Wong
Y H
YH
D037361:Adenylate Cyclase Toxin; D000067956:Adenylyl Cyclase Inhibitors; D002460:Cell Line; D005576:Colforsin / Q000494:pharmacology; D000242:Cyclic AMP / Q000037:antagonists & inhibitors*; D004305:Dose-Response Relationship, Drug; D019204:GTP-Binding Proteins / Q000378:metabolism; D006801:Humans; D007074:Immunoglobulin G / Q000235:genetics; D018847:Opioid Peptides / Q000494:pharmacology; D037342:Pertussis Toxin; D011961:Receptors, Fc / Q000235:genetics; D011957:Receptors, Opioid / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D016324:Sequence Tagged Sites; D010566:Virulence Factors, Bordetella / Q000494:pharmacology; D000094942:Nociceptin Receptor; D000097629:Nociceptin
D037361:Adenylate Cyclase Toxin; D000067956:Adenylyl Cyclase Inhibitors; D005576:Colforsin; D000242:Cyclic AMP; D019204:GTP-Binding Proteins; D007074:Immunoglobulin G; D018847:Opioid Peptides; D037342:Pertussis Toxin; D011961:Receptors, Fc; D011957:Receptors, Opioid; D010566:Virulence Factors, Bordetella; D000094942:Nociceptin Receptor; D000097629:Nociceptin; C000722283:OPRL1 protein, human