Ozaki Yuichi, Fukada Haruhisa, Kazeto Yukinori, Adachi Shinji, Hara Akihiko, Yamauchi Kohei
Division of Marine Biosciences, Graduate School of Fisheries Sciences, Hokkaido University, Hakodate, Hokkaido 041-8611, Japan.
Comp Biochem Physiol B Biochem Mol Biol. 2006 Apr;143(4):422-31. doi: 10.1016/j.cbpb.2005.12.016. Epub 2006 Feb 3.
Two cDNAs encoding growth hormone receptor (GHR)-like genes, eGHR1 and eGHR2, were isolated from Japanese eel (Anguilla japonica) liver tissue. The putative eel GHR proteins showed conserved structural features of vertebrate GHRs, including six cysteine residues and a YGEFS motif in the extracellular domain, a single transmembrane region, and proline-rich box 1 and box 2 domains. Northern blot analysis showed a single eGHR1 transcript in liver, while two sizes of eGHR2 transcripts, thought to be produced by alternative splicing, were present. RT-PCR revealed that eGHR1 and eGHR2 transcripts were widely distributed throughout the whole body of the Japanese eel. Moreover, the results of binding assays showed the specific binding of growth hormone to recombinant eGHR1. Since these putative eGHR proteins show all characteristics of the GHR family, we conclude that eGHR1 and eGHR2 cDNA encode two different GHRs in Japanese eel. We confirmed the ligand specificity of eGHR1 by binding assay, and further research is needed to allow characterization of the binding capability of eGHR2.
从日本鳗鲡(Anguilla japonica)肝脏组织中分离出两个编码类生长激素受体(GHR)基因的cDNA,即eGHR1和eGHR2。推测的鳗鲡GHR蛋白具有脊椎动物GHR的保守结构特征,包括细胞外结构域中的六个半胱氨酸残基和一个YGEFS基序、一个单一跨膜区域以及富含脯氨酸的框1和框2结构域。Northern印迹分析显示肝脏中有一个单一的eGHR1转录本,而存在两种大小的eGHR2转录本,推测是由可变剪接产生的。逆转录聚合酶链反应(RT-PCR)表明,eGHR1和eGHR2转录本在日本鳗鲡的全身广泛分布。此外,结合试验结果显示生长激素与重组eGHR1有特异性结合。由于这些推测的eGHR蛋白具有GHR家族的所有特征,我们得出结论,eGHR1和eGHR2 cDNA在日本鳗鲡中编码两种不同的GHR。我们通过结合试验证实了eGHR1的配体特异性,还需要进一步研究以确定eGHR2的结合能力。