MOLECULAR-CLONING OF A GENE ENCODING THE HISTAMINE-H2-RECEPTOR
MOLECULAR-CLONING OF A GENE ENCODING THE HISTAMINE-H2-RECEPTOR
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DOI：
10.1073/pnas.88.2.429
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发表时间：
1991-01-01
影响因子：
11.1
通讯作者：
YAMADA, T
中科院分区：
文献类型：
--
作者：
GANTZ, I;SCHAFFER, M;YAMADA, T
组胺受体的H2亚类介导胃酸分泌，该受体的拮抗剂已被证明是胃肠道酸性消化性疾病的有效疗法。 组胺的生理作用已被证明是通过与腺苷酸环化酶激活和细胞cAMP生成相关的鸟嘌呤核苷酸结合蛋白介导的。 我们利用聚合酶链反应技术，使用简并寡核苷酸引物的基础上连接到鸟嘌呤核苷酸结合蛋白的细胞受体之间的已知同源性，以获得犬胃壁细胞cDNA的部分长度克隆。 该克隆用于从犬基因组文库获得全长受体基因。 组胺以剂量依赖性方式增加细胞cAMP含量在L细胞永久转染该基因，和预孵育的细胞与H2-选择性拮抗剂西咪替丁转移的剂量-反应曲线的权利。 西咪替丁以剂量依赖性方式抑制放射性标记的H2受体选择性配体[甲基-H-3]噻替丁与转染细胞的结合，但H1选择性拮抗剂苯海拉明则不然。 这些数据表明，我们已经克隆了一个基因，编码H2亚型的组胺受体。
The H2 subclass of histamine receptors mediates gastric acid secretion, and antagonists for this receptor have proven to be effective therapy for acid peptic disorders of the gastrointestinal tract. The physiological action of histamine has been shown to be mediated via a guanine nucleotide-binding protein linked to adenylate cyclase activation and cellular cAMP generation. We capitalized on the technique of polymerase chain reaction, using degenerate oligonucleotide primers based on the known homology between cellular receptors linked to guanine nucleotide-binding proteins to obtain a partial-length clone from canine gastric parietal cell cDNA. This clone was used to obtain a full-length receptor gene from a canine genomic library. Histamine increased in a dose-dependent manner cellular cAMP content in L cells permanently transfected with this gene, and preincubation of the cells with the H2-selective antagonist cimetidine shifted the dose-response curve to the right. Cimetidine inhibited the binding of the radiolabeled H2 receptor-selective ligand [methyl-H-3]tiotidine to the transfected cells in a dose-dependent fashion, but the H1-selective antagonist diphenhydramine did not. These data indicate that we have cloned a gene that encodes the H2 subclass of histamine receptors.