克隆人类κ阿片受体的大脑。
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陈竺J, C,雪JC, Kunapuli年代,DeRiel JK,刘晨LY
克隆人类κ阿片受体的大脑。
生命科学。1995;56 (9):PL201-7。
- PubMed ID
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7869844 (在PubMed]
- 文摘
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通过使用一个老鼠κ阿片受体cDNA探针筛选一个人类大脑cDNA文库,我们孤立4.0 kb克隆(z115)包括一个主要部分,人类κ阿片受体(hkor),从氨基酸残基# 6 - 3 '非翻译区。极端的5 '地区232 - bp z115碎片被用作探针筛选一个人类基因组DNA库。1.6 kb片段(d2)的一个积极的克隆被发现从5 '非翻译区扩展到超出了外显子/内含子结渣Arg86。基因组DNA片段d2和cDNA克隆z115被组装生成一个克隆(d2-z115)包含整个hkor编码序列。克隆d2-z115 1140个基点的开放阅读框,编码380个氨基酸蛋白质。推导的氨基酸序列有93.9%和93.2%的身份老鼠和老鼠κ受体,分别。它还显示大约60%的身份人类μ和δ受体。北部污点分析表明,在人类的大脑有一个hkor mRNA转录6.0 kb。大脑区域检查、杏仁核、尾状核、下丘脑和丘脑核含有高水平的hkor mRNA。Hkor被克隆到表达载体pBK-CMV和瞬变COS-1细胞中表达。 Hkor had high affinity for [3H] diprenorphine, a nonselective opioid antagonist, and displayed stereospecific binding to naloxone. kappa selective ligands (U50,488H and nor-BNI) had high affinities, whereas mu and delta selective ligands bound with much lower affinities. Dynorphin A (1-17) and alpha-neoendorphin, both endogenous kappa peptides, bound with high affinities. These binding characteristics confirmed that hkor is a kappa receptor, most likely kappa 1 type. Cloning of the human kappa receptor allows investigation of interactions of compounds with the human receptor, instead of rodent receptors, for development of better therapeutic agents.