Opioid receptors from a lower vertebrate (Catostomus commersoni): Sequence, pharmacology, coupling to a G-protein-gated inward-rectifying potassium channel (GIRK1), and evolution

被引:63
作者
Darlison, MG
Greten, FR
Harvey, RJ
Kreienkamp, HJ
Stuhmer, T
Zwiers, H
Lederis, K
Richter, D
机构
[1] UNIV HAMBURG,INST ZELLBIOCHEM & KLIN NEUROBIOL,D-20246 HAMBURG,GERMANY
[2] UNIV CALGARY,HLTH SCI CTR,DEPT PHYSIOL & BIOPHYS,CALGARY,AB T2N 4N1,CANADA
[3] UNIV CALGARY,HLTH SCI CTR,DEPT THERAPEUT & PHARMACOL,CALGARY,AB T2N 4N1,CANADA
关键词
cDNA cloning; functional expression; peptide receptor; teleost fish; Xenopus oocyte;
D O I
10.1073/pnas.94.15.8214
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
The molecular evolution of the opioid receptor family has been studied by isolating cDNAs that encode six distinct opioid receptor-like proteins from a lower vertebrate, the teleost fish Catostomus commersoni. One of these, which has been obtained in full-length form, encodes a 383-amino acid protein that exhibits greatest sequence similarity to mammalian mu-opioid receptors; the corresponding gene is expressed predominantly in brain and pituitary, Transfection of the teleost cDNA into HEK 293 cells resulted in the appearance of a receptor having high affinity for the mu-selective agonist [D-Ala(2), MePhe(4)-Gly-ol(5)] enkephalin (DAMGO) (K-d = 0.63 +/- 0.15 nM) and for the nonselective antagonist naloxone (K-d = 3.1 +/- 1.3 nM), The receptor had negligible affinity for U50488 and [D-Pen(2), D-Pen(5)]enkephalin (DP-DPE), which are kappa- and delta-opioid receptor selective agonists, respectively. Stimulation of transfected cells with 1 mu M DAMGO lowered forskolin-induced cAMP levels, an effect that could be reversed by naloxone, Experiments in Xenopus oocytes have demonstrated that the fish opioid receptor can, in an agonist-dependent fashion, activate a coexpressed mouse G-protein-gated inward-rectifying potassium channel (GIRK1), The identification of six distinct fish opioid receptor-like proteins suggests that additional mammalian opioid receptors remain to be identified at the molecular level, Furthermore, our data indicate that the mu-opioid receptor arose very early in evolution, perhaps before the appearance of vertebrates; and that the pharmacological and functional properties of this receptor have been conserved over a period of approximate to 400 million years implying that it fulfills an important physiological role.
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页码:8214 / 8219
页数:6
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