グアニリンの分子構造、細胞学、生理学とその臨床応用

鸟苷酸的分子结构、细胞学、生理学及其临床应用

• 批准号: 06044104
• 负责人: HAYAKAWA Tetsuo
• 金额: $ 7.74万
• 依托单位: Nagoya University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for international Scientific Research
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-06044104/
• 关键词: guanylin; proguanylin; immunohistochemistry; gastrointestinal tract; kidney; radioreceptor assay; molecular structure; autoradiography; (1)グアニリン; (2)プログアニリン; (3)免疫組織化学; (4)消化管; (5)腎臓; (6)ラジオレセプターアッセイ; (7)分子構造; (8)オートラジオグラフィー; エンテロトキシン; 異性

Guanylin, isolated from rat small intestine, is a 15 amino acid residue peptide structurally related to the heat-stable enterotoxin (STa) of E.coli. In order to understand its function in health and disease states, we have synthesized guanylin and its precursor related peptides and tried to determine their structure and bindings to their receptors and to identify its producing cells and their tharget organs. Nuclear magnetic resonance spectroscopy revealed that guanylin with disulfids positions 4-12 and 7-15 exists as a mixture of two stable conformations with compact spiral structures. Its isomer with disulfides positions 4-15 and 7-12 showed a single conformation.Immunohistochemical studies in human and rat intestine using antiserum against proguanylin 1-15 indicated that proguanylin-like immunoreactivities were present in the intestinal endocrine cells which were also positive somatostatin but negative to serotonin, suggesting proguanyline may be cosecreted with somatostatin. Progua … More nyline positive cells were abundant in the pyrolic antrum and duodenum. Very few cells were observed in the colon. Both guanylin and STa displaced the binding of ^<125>I-labelled guanylin to crude membrane preparations of rat small intestine in a dose dependent manner with Kd values around 10^<-6> M.The disulfide isomer, on the other hand, bound poorly to the membrane preparations. Most of ^<125>I-guanylin was incorporated into the kidney following the intravenous injection in anesthetized rats. It bound to the luminal surface of proximal tubules. In the collecting ducts, ^<125>I-guanylin appeared to be excreted via chief cells. Guanylin decreased the cell height and increased the luminal space of the inner medullary collecting ducts, suggesting that guanylin has a diuretic action in the collecting ducts. In the small intestine guanylin stimulated mucus secretion from goblet cells in crypts but not in the villi. It appears that guanylin control both intestinal and renal fluid secretion as both paracrine and endocrine hormones. Less

鸟苷素是从大鼠小肠中分离的一种15个氨基酸残基的肽，其结构与大肠杆菌的热稳定肠毒素（STa）相关。为了了解鸟苷素在健康和疾病状态下的功能，我们合成了鸟苷素及其前体相关肽，并试图确定它们的结构和与其受体的结合，并鉴定其产生细胞及其靶器官。核磁共振光谱显示，鸟苷素与二硫化物的位置4-12和7-15存在作为两个稳定的构象与紧凑的螺旋结构的混合物。用抗原鸟苷素1 -15的抗血清对人和大鼠小肠进行免疫组化研究，发现原鸟苷素样免疫反应存在于肠内分泌细胞中，这些细胞也是生长抑素阳性细胞，但对5-羟色胺阴性，提示原鸟苷素可能与生长抑素共同分泌。普罗瓜 ...更多信息 在胃窦和十二指肠内有大量的nyline阳性细胞。在结肠中观察到非常少的细胞。鸟苷素和STa都<125>以剂量依赖性方式取代了~ 13 I标记的鸟苷素与大鼠小肠粗膜制剂的结合，Kd值约为10 μ <-6>M。另一方面，二硫键异构体与膜制剂的结合较差。在<125>麻醉大鼠中静脉注射后，大部分^I-鸟苷素掺入肾脏。它结合到近端小管的管腔表面。在集合管中，^<125>I-鸟苷素似乎通过主细胞排出。鸟苷素降低了细胞高度，增加了内髓集合管的管腔空间，表明鸟苷素在集合管中具有利尿作用。在小肠中，鸟苷素刺激隐窝中的杯状细胞分泌粘液，但不刺激绒毛中的杯状细胞。鸟苷素作为旁分泌激素和内分泌激素控制肠和肾的液体分泌。少

项目成果

H.Ieda,et al.: "Localization of proguanylin examined by its Nterminal specific antisera in the rat gastrointestinal tract" 解剖学雑誌. 71. 396 (1996)

H.Ieda 等人：“通过其 N 末端特异性抗血清在大鼠胃肠道中检查原鸟苷蛋白的定位”《解剖学杂志》71. 396 (1996)。

K Nokihara,et al.: "Two-dimensional electrophoresis as a complementary method of isolating peptide fragments of cleaved proteins for internal sequencing" J.Chorm.676. 233-238 (1994)

K Nokihara 等人：“二维电泳作为分离裂解蛋白肽片段以进行内部测序的补充方法”J.Chorm.676。

家田秀明、他: "消化管におけるプログアニリン産生細胞の局在" 日本消化器病学会雑誌. 92. 1539 (1995)

Hideaki Ieda 等：“胃肠道中原鸟苷蛋白产生细胞的定位”日本胃肠病学会杂志 92. 1539 (1995)。

K Nokihara,et al.: "Highly efficient peptide-smide preparation using a simultaneous multiple-peptide synthesizer with two novel acid labile linkers" Peptides:Chemistry and Biology. 85-87 (1994)

K Nokihara 等人：“使用具有两个新型酸不稳定接头的同步多肽合成器进行高效肽-smide 制备”肽：化学和生物学。

K Nokihara,et al.: "Chemical synthesis of pro-guanylin related peptides for biochemical and immunochemical studies." Peptide Chemisty 1995. 117-120 (1996)

K Nokihara 等人：“用于生化和免疫化学研究的鸟苷蛋白原相关肽的化学合成。”