HUMAN SUBSTANCE-P RECEPTOR (NK-1) - ORGANIZATION OF THE GENE, CHROMOSOME LOCALIZATION, AND FUNCTIONAL EXPRESSION OF CDNA CLONES
HUMAN SUBSTANCE-P RECEPTOR (NK-1) - ORGANIZATION OF THE GENE, CHROMOSOME LOCALIZATION, AND FUNCTIONAL EXPRESSION OF CDNA CLONES
复制标题
DOI：
10.1021/bi00108a006
复制
发表时间：
1991-11-05
期刊：
影响因子：
2.9
通讯作者：
GERARD, C
中科院分区：
文献类型：
--
作者：
GERARD, NP;GARRAWAY, LA;GERARD, C
利用引物聚合酶链反应制备cDNA探针，根据大鼠序列克隆人P物质受体（NK-1）基因。该基因全长45- 60kb，包含在5个外显子中，内含子在与NK-2受体基因同源的位点中断。对小鼠/人类杂交细胞基因组DNA的限制性酶切分析表明，NK-1受体是位于人类2号染色体上的单拷贝基因。利用基于编码序列5′和3′端引物的聚合酶链反应，从人肺和IM9淋巴母细胞中生成全长cdna。转染COS-7细胞后，NK-1受体结合I-125-BHSP， K(d)为0.35 +/- 0.07 nM，介导P物质诱导的磷脂酰肌醇代谢。受体对P物质有选择性；对神经激肽A和神经激肽B的相对亲和力分别低100倍和500倍。人IM9淋巴母细胞表达相对较高水平的NK-1受体，Northern blot分析表明糖皮质激素和生长因子可调节mRNA水平，提示该细胞系可作为研究NK-1受体基因表达控制的模型。
The gene for the human substance P receptor (NK-1) was cloned using cDNA probes made by the polymerase chain reaction from primers based on the rat sequence. The gene spans 45-60 kb and is contained in five exons, with introns interrupting at sites homologous to those in the NK-2 receptor gene. Analysis of restriction digests of genomic DNA from mouse/human cell hybrids indicates the NK-1 receptor is a single-copy gene located on human chromosome 2. Polymerase chain reaction using primers based on the 5' and 3' ends of the coding sequence was used to generate full-length cDNAs from human lung and from IM9 lymphoblast cells. When transfected into COS-7 cells, the NK-1 receptor binds I-125-BHSP with a K(d) of 0.35 +/- 0.07 nM and mediates substance P induced phosphatidylinositol metabolism. The receptor is selective for substance P; the relative affinity for neurokinin A and neurokinin B is 100- and 500-fold lower, respectively. Human IM9 lymphoblast cells express relatively high levels of the NK-1 receptor, and Northern blot analysis indicates modulation of mRNA levels by glucocorticoids and growth factors, suggesting that this cell line may be useful as a model for studying the control of NK-1 receptor gene expression.