Habert-Ortoli E, Amiranoff B, Loquet I, Laburthe M, Mayaux J F
Rhône-Poulenc Rorer SA, Centre de Recherche de Vitry-Alfortville, Département Biotechnologie, Vitry sur Seine, France.
Proc Natl Acad Sci U S A. 1994 Oct 11;91(21):9780-3. doi: 10.1073/pnas.91.21.9780.
The ubiquitous neuropeptide galanin controls numerous functions such as endocrine secretions, intestinal motility, and behavioral activities. These regulatory effects of galanin are mediated through the interaction with specific membrane receptors and involve the pertussis toxin-sensitive guanine nucleotide binding proteins Gi/Go as transducing elements. We report here the isolation of a cDNA coding for a human galanin receptor from a Bowes melanoma cell line cDNA expression library, by using a radioligand binding strategy. The nucleotide sequence of the cloned receptor reveals an open reading frame encoding a 349-amino acid protein with seven putative hydrophobic transmembrane domains and significant homology with members of the guanine nucleotide binding protein-coupled neuropeptide receptor family. The cloned receptor expressed in COS cells specifically binds human, porcine, and rat galanin with high affinity (Kd in the nanomolar range) and mediates the galanin inhibition of adenylate cyclase. A 2.8-kb galanin receptor transcript was identified in several human tissues. Cloning of this galanin receptor should enhance our knowledge of its distribution, structure, and function in human physiology and pathophysiology.
普遍存在的神经肽甘丙肽可控制多种功能,如内分泌分泌、肠道蠕动和行为活动。甘丙肽的这些调节作用是通过与特定膜受体相互作用介导的,并且涉及百日咳毒素敏感的鸟嘌呤核苷酸结合蛋白Gi/Go作为转导元件。我们在此报告,通过使用放射性配体结合策略,从鲍斯黑色素瘤细胞系cDNA表达文库中分离出编码人甘丙肽受体的cDNA。克隆受体的核苷酸序列揭示了一个开放阅读框,编码一个349个氨基酸的蛋白质,该蛋白质具有七个推定的疏水跨膜结构域,并且与鸟嘌呤核苷酸结合蛋白偶联的神经肽受体家族成员具有显著同源性。在COS细胞中表达的克隆受体以高亲和力(Kd在纳摩尔范围内)特异性结合人、猪和大鼠甘丙肽,并介导甘丙肽对腺苷酸环化酶的抑制作用。在几种人体组织中鉴定出一个2.8kb的甘丙肽受体转录本。克隆这种甘丙肽受体应能增强我们对其在人体生理学和病理生理学中的分布、结构和功能的了解。