Luebke A E, Dahl G P, Roos B A, Dickerson I M
Department of Physiology and Biophysics, Veterans Affairs Medical Center, University of Miami School of Medicine, FL 33101, USA.
Proc Natl Acad Sci U S A. 1996 Apr 16;93(8):3455-60. doi: 10.1073/pnas.93.8.3455.
An expression-cloning strategy was used to isolate a cDNA that encodes a protein that confers calcitonin gene-related peptide (CGRP) responsiveness to Xenopus laevis oocytes. A guinea pig organ of Corti (the mammalian hearing organ) cDNA library was screened by using an assay based on the cystic fibrosis transmembrane conductance regulator (CFTR). The CFTR is a chloride channel that is activated upon phosphorylation; this channel activity was used as a sensor for CGRP-induced activation of intracellular kinases. A cDNA library from guinea pig organ of Corti was screened by using this oocyte-CFTR assay. A cDNA was identified that contained an open reading frame coding for a small hydrophilic protein that is presumed to be either a CGRP receptor or a component of a CGRP receptor complex. This CGRP receptor component protein confers CGRP-specific activation to the CFTR assay, as no activation was detected upon application of calcitonin, amylin, neuropeptide Y, vasoactive intestinal peptide, or beta-endorphin. In situ hybridization demonstrated that the CGRP receptor component protein is expressed in outer hair cells of the organ of Corti and is colocalized with CGRP-containing efferent nerve terminals.
采用表达克隆策略分离出一个编码赋予非洲爪蟾卵母细胞降钙素基因相关肽（CGRP）反应性蛋白的cDNA。利用基于囊性纤维化跨膜传导调节因子（CFTR）的检测方法筛选豚鼠柯蒂氏器（哺乳动物听觉器官）cDNA文库。CFTR是一种经磷酸化后被激活的氯离子通道；该通道活性被用作CGRP诱导的细胞内激酶激活的传感器。通过这种卵母细胞 - CFTR检测方法筛选豚鼠柯蒂氏器的cDNA文库。鉴定出一个cDNA，其包含一个编码小亲水性蛋白的开放阅读框，该蛋白推测为CGRP受体或CGRP受体复合物的一个组分。这种CGRP受体组分蛋白赋予CFTR检测方法CGRP特异性激活，因为在应用降钙素、胰淀素、神经肽Y、血管活性肠肽或β - 内啡肽时未检测到激活。原位杂交表明，CGRP受体组分蛋白在柯蒂氏器的外毛细胞中表达，并与含CGRP的传出神经末梢共定位。
