Study on recognition mechanism of heat-stable enterotoxin by its receptorprotein
热稳定肠毒素受体蛋白识别机制研究
基本信息
- 批准号:04680160
- 负责人:
- 金额:$ 1.22万
- 依托单位:
- 依托单位国家:日本
- 项目类别:Grant-in-Aid for General Scientific Research (C)
- 财政年份:1992
- 资助国家:日本
- 起止时间:1992 至 1993
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
Heat-stable enterotoxins (STs) are peptides of 18 or 19 amino acid residues produced by enterotoxigenic Escherichia coli and are known to cause acute diarrhea in men and animals. The biological action of ST is initiated by the binding to its receptor protein on the membrane of intestinal epithelial cell. The formation of a ST-receptor complex leads to stimulation of guanylate cyclase and is followed by increase in GMP concentration in the cells and fluid secretion from the cells. This study is planned to identify the binding part of the receptor protein in its binding to ST, purify and crystallize the complex j of ST with its receptor protein for its X-ray crystallographic analysis in order to elucidate the co-recogniton mechanism between ST and its receptor protein which is the first step of signal transduction by ST via the receptor protein on the membrane of cells. To accomplish this purpose, I synthesized a variety of ST analogues were labeled with the fluorophore and cific binding abilties of these ST analogues were examined by means of chromatographic method. Among these, there were two ST analogues with the specific binding ability to the receptor protein corresponding to that of the native ST.One of these analogues was used to form the complex with the receptor protein on the membrane prepared from rat intestine. After solubilization experimental conditions to purify the complex were examined and the conbination of several kinds of HPLC lead to obtain a partially purified complex with fluorescence to get the knowledge about the binding part of receptor protein to ST, the purification of the protein crosslinked with ST labeled with the fluorophore is in progress.
热稳定性肠毒素(STs)是由产肠毒素的大肠杆菌产生的18或19个氨基酸残基的肽,已知可引起人类和动物的急性腹泻。ST的生物学作用是通过与肠上皮细胞膜上的受体蛋白结合而启动的。st受体复合物的形成导致鸟苷酸环化酶的刺激,随后是细胞内GMP浓度的增加和细胞分泌液的分泌。本研究拟鉴定受体蛋白与ST结合的结合部位,纯化并结晶ST与其受体蛋白的复合物j进行x射线晶体学分析,以阐明ST与其受体蛋白的共识别机制,这是ST通过细胞膜上受体蛋白进行信号转导的第一步。为了达到这个目的,我合成了多种ST类似物并用荧光团标记,并用色谱法检测了这些ST类似物的特异性结合能力。其中,有两种ST类似物与受体蛋白的特异性结合能力与天然ST相对应,其中一种类似物与受体蛋白在制备的大鼠肠膜上形成复合物。在考察了增溶实验条件纯化配合物后,结合几种高效液相色谱法得到部分纯化的带荧光的配合物,以了解受体蛋白与ST的结合部分,与ST交联的蛋白用荧光团标记的纯化正在进行中。
项目成果
期刊论文数量(4)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Takehiro Okumura, Takashi Sato, Hiroshi Ozaki, Akihiro Wada, Yuji Hidaka, and Yasutsugu Shimonishi: "Structure-Function Relationship of Heat-Stable Enterotoxin (ST) : Steric Requirments at Position 12 for Receptor Binding" Peptide Chemistry. 217-220 (1994
Takehiro Okumura、Takashi Sato、Hiroshi Ozaki、Akihiro Wada、Yuji Hidaka 和 Yasutsugu Shimonishi:“热稳定肠毒素 (ST) 的结构-功能关系:受体结合的 12 位空间要求”肽化学。
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- 影响因子:0
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- 通讯作者:
Takehiro Okumura,Takashi Sato,Hiroshi Ozaki,Akihiro Wada,Yuji Hidaka,and Yasutsugu Shimonishi: "Structure-Function Relationship of Heat-Stable Enterotoxin(ST):Steric Requirments at Position 12 for Receptor Binding" Peptide Chemistry 1993. 217-220 (1994)
Takehiro Okumura、Takashi Sato、Hiroshi Ozaki、Akihiro Wada、Yuji Hidaka 和 Yasutsugu Shimonishi:“热稳定肠毒素 (ST) 的结构-功能关系:受体结合的位置 12 的空间要求”肽化学 1993. 217-220 (
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OZAKI Hiroshi其他文献
OZAKI Hiroshi的其他文献
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{{ truncateString('OZAKI Hiroshi', 18)}}的其他基金
Studies on the phased immune-barrier systems in gut-liver axis focusing on immune responses of mesenchymal cells
肠肝轴阶段性免疫屏障系统研究,重点关注间充质细胞的免疫反应
- 批准号:
20228005 - 财政年份:2008
- 资助金额:
$ 1.22万 - 项目类别:
Grant-in-Aid for Scientific Research (S)
Immunological approach for the molecular mechanism ofintestinal motility functions
肠蠕动功能分子机制的免疫学方法
- 批准号:
16208029 - 财政年份:2004
- 资助金额:
$ 1.22万 - 项目类别:
Grant-in-Aid for Scientific Research (A)
Clarification of new cell signaling mediated by intermediairy metabolite of claolesterol
阐明长甾醇中间代谢物介导的新细胞信号传导
- 批准号:
14360176 - 财政年份:2002
- 资助金额:
$ 1.22万 - 项目类别:
Grant-in-Aid for Scientific Research (B)
Intestinal motility immune systems in Hirschsprung's disease model animals
先天性巨结肠模型动物的肠道运动免疫系统
- 批准号:
12460132 - 财政年份:2000
- 资助金额:
$ 1.22万 - 项目类别:
Grant-in-Aid for Scientific Research (B)
Studies on novel marine bioactive substances as pharmacological resources
新型海洋生物活性物质药理资源研究
- 批准号:
10356011 - 财政年份:1998
- 资助金额:
$ 1.22万 - 项目类别:
Grant-in-Aid for Scientific Research (A)
Organculture as a new model of atherosclerosis
器官培养作为动脉粥样硬化的新模式
- 批准号:
09660315 - 财政年份:1997
- 资助金额:
$ 1.22万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
Structure and function of proteins to form complex with receptor for heat-stable enterotoxin
与热稳定肠毒素受体形成复合物的蛋白质的结构和功能
- 批准号:
06680583 - 财政年份:1994
- 资助金额:
$ 1.22万 - 项目类别:
Grant-in-Aid for General Scientific Research (C)
Endothelin receptor subtype and physiology of vascular smooth muscle and endothelium
内皮素受体亚型与血管平滑肌和内皮的生理学
- 批准号:
04454115 - 财政年份:1992
- 资助金额:
$ 1.22万 - 项目类别:
Grant-in-Aid for General Scientific Research (B)
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