Comparative molecular biological studies on the vasoactive peptide receptors.

血管活性肽受体的比较分子生物学研究。

• 批准号: 04404082
• 负责人: HIROSE Shigehisa
• 金额: $ 14.08万
• 依托单位: Tokyo Institute of Technology
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (A)
• 财政年份: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-04404082/
• 关键词: natriuretic peptide; receptor; subtype; endothelin; chondrocyte; cloning; oligopeptidase; オリゴペプチダーゼ; SABP; 点突然変異体; ジスルフィド結合; 免疫組織化学染色; 内皮細胞; サブタイプ; 情報伝達系; 択一的RNAスプライシング; 分子生物学; ウナギ; 軟骨組織

To further advance our understanding on the physiological significance of vasoactive peptides, we carried out the following comparative molecular biological studies on their receptors.1) Natriuretic peptide receptors : The intra-and inter-subunit disulfide bonding pattern of the bovine and eel natriuretic peptide receptor C (NPR-C) were determined by a combination of protein sequencing and site-directed mutagenesis. Analysis of the cDNA and gene structures of mammalian NPR-C revealed the presence of an alternatively spliced mRNA species that encodes a receptor protein with a slightly modified sequence in the juxta-membrane region of the extracellular domain. A new receptor subtype, termed NPR-D,has been identified, cloned, and characterized. NPR-D has a short cytoplasmic tail and is 70% similar to the NPR-C sequence, but quite different in the tissue distribution ; NPR-D is abundant in the brain.2)Endothelin receptors : Localization of the endothelin receptor B (ET-B) was determined by immunohistochemistry and the vascular endotherium was found to be decorated most densely. Type A endothelin receptor (ET-A) was found to be highly expressed in the perichondrium of rats and the gill epitherium of trout, suggesting that endothelin is involved in the differentiation of chondrocytes and regulation of gill functions.

为了进一步加深对血管活性肽生理意义的理解，我们对其受体进行了以下比较分子生物学研究。1)利尿钠肽受体：牛和鳗鱼利钠肽受体的亚基内和亚基间二硫键键合模式C (NPR-C) 通过蛋白质测序和定点诱变相结合来确定。对哺乳动物 NPR-C 的 cDNA 和基因结构的分析揭示了存在一种可变剪接的 mRNA 种类，该 mRNA 编码的受体蛋白在胞外结构域的近膜区域中序列略有修改。一种称为 NPR-D 的新受体亚型已被鉴定、克隆和表征。 NPR-D胞浆尾短，与NPR-C序列70%相似，但组织分布差异较大； NPR-D在大脑中含量丰富。2）内皮素受体：通过免疫组织化学测定内皮素受体B（ET-B）的定位，发现血管内皮装饰最密集。 A型内皮素受体（ET-A）在大鼠软骨膜和鳟鱼鳃上皮中高表达，提示内皮素参与软骨细胞分化和鳃功能调节。

项目成果

Makoto Itakura: "Mutational Analysis of Disulfide Bridges in the Type C Atrial Natriuretic Peptid Receptor" Journal of Biological Chemistry. (in press).

Makoto Itakura：“C 型心房钠尿肽受体中二硫键的突变分析”生物化学杂志。

Hiromi Hagiwara, et.al.: "Natriuretic peptides and their receptors." Zool.Sci.12. 141-149 (1995)

Hiromi Hagiwara 等人：“钠尿肽及其受体。”

Masatora Iwashina: "His^<145>-Trp^<146> Residues and the Disulfide-Linked Loops in Atrial Natriuretic Peptide Receptor Are Critical for the Ligand-Binding Activity" Journal of Biological Chemistry. (in press).

Masatora Iwashina：“心房钠尿肽受体中的His^<145>-Trp^<146>残基和二硫键连接环对于配体结合活性至关重要”《生物化学杂志》。

Takeshi Katafuchi: "A dramatic pH-dependent alteration in ANP receptor density : A note for using cultured cells." Americal Journal of Physiology. 264. C1345-C1349 (1993)

Takeshi Katafuchi：“ANP 受体密度的 pH 依赖性显着变化：使用培养细胞的注意事项。”

Akira Kato, et.al.: "Similarities of 'soluble angiotensin-binding protein" and 'liver microsomal endopeptidase" suggest that they are identical and represent a new member of the thimet oligopeptidase family which includes metal leoendopeptidase 24 15." Eu

Akira Kato 等人：“‘可溶性血管紧张素结合蛋白’和‘肝微粒体肽链内切酶’的相似性表明它们是相同的，并且代表了 thimet 寡肽酶家族的新成员，该家族包括金属 leo 肽链内切酶 24 15。”