Molecular cloning and pharmacological characterization of a somatostatin receptor subtype in the gymnotiform fish Apteronotus albifrons

被引:31
作者
Zupanc, GKH
Siehler, S
Jones, EMC
Seuwen, K
Furuta, H
Hoyer, D
Yano, H
机构
[1] Univ Manchester, Sch Biol Sci, Manchester M13 9PT, Lancs, England
[2] Univ Chicago, Howard Hughes Med Inst, Dept Biochem & Mol Biol, Chicago, IL 60637 USA
[3] Univ Chicago, Howard Hughes Med Inst, Dept Med, Chicago, IL 60637 USA
[4] Max Planck Inst Dev Biol, Dept Phys Biol, D-72011 Tubingen, Germany
[5] Univ Manchester, Sch Biol Sci, Manchester M13 9PT, Lancs, England
[6] Novartis Pharma AG, Nervous Syst Res, CH-4002 Basel, Switzerland
关键词
D O I
10.1006/gcen.1999.7316
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
The actions of the various forms of somatostatin (SRIF), including those of the tetradecapeptide SRIF14, are mediated by specific receptors. In mammals, five subtypes of SRIF receptors, termed sst(1-5), have been cloned. Using a combination of reverse transcriptase-polymerase chain reaction and genomic library screening in the gymnotiform fish Apteronotus albifrons, a gene encoding the first-known nonmammalian SRIF receptor has been isolated, The deduced amino acid sequence displays 59% identity with the human sst(3) receptor protein; hence, the gene is termed "Apteronotus sst(3)." The predicted protein consists of 494 amino acid residues exhibiting a putative seven-transmembrane domain topology typical of G protein-coupled receptors. A signal corresponding to the Apteronotus sst(3) receptor was detected in brain after amplification of poly(A)(+)-RNA by reverse transcriptase-polymerase chain reaction, but not by Northern blot analysis or in situ hybridization, suggesting a low level of expression. Membranes prepared from CCL39 cells stably expressing the Apteronotus sst(3) receptor gene bound [I-125][Leu(8),D-Trp(22),(125) I-Tyr(25)]SRIF28 with high affinity and in a saturable manner (B-max = 4470 fmol/mg protein; pK(D) = 10.5). SRIF14 and various synthetic SRIF receptor agonists produced a dose-dependent inhibition of radioligand binding, with the following rank order of potency: SRIF14 approximate to SRIF28 > BIM 23052 > octreotide > BIM 23056. Under low stringency conditions, an Apteronotus sst(3) probe hybridized to multiple DNA fragments in HindIII or EcoRI digests of A. albifrons DNA, indicating that the Apteronotus sst(3) receptor is a member of a larger family of Apteronotus SRIF receptors. (C) 1999 Academic Press.
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页码:333 / 345
页数:13
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