课题基金 / 基金详情

Interaction of multiple phospholipase C and GTP-binding proteins in platelet signal transduction

Interaction of multiple phospholipase C and GTP-binding proteins in platelet signal transduction
多种磷脂酶 C 和 GTP 结合蛋白在血小板信号转导中的相互作用
批准号:
02454544
负责人:
NOZAWA Yoshinori
金额:
$4.35万
依托单位:
依托单位国家:
日本
项目类别:
Grant-in-Aid for General Scientific Research (B)
财政年份:
1990
资助国家:
日本
项目状态:
已结题
起止时间:
1990 至 1992

项目摘要

项目成果

NOZAWA Yoshinori的其他基金

相似基金

相关文献

中文摘要
翻译
众所周知,激动剂-受体相互作用后对磷脂酰肌醇周转的刺激是通过质膜上假定的GTP结合蛋白激活磷脂酰肌醇特异性磷脂酶C(PI-PLC)。血小板常被用作研究跨膜信号的一个潜在有用的模型。本研究的目的是通过纯化PI-PLC同工酶和GTP结合蛋白,并充分结合重组,阐明PI-PLC在人血小板中激活的机制。对人血小板胞浆和膜组分的PI-PLC的有效拆分显示出五个明显的活性峰。用不同的PI-PLC同工酶抗体进行Western blotting分析,结果表明,PLC-β、PLC-γ和PLC-Delta同工酶被鉴定出来,并从胞浆组分中分离出另外两个未知的活性峰,PLC-β主要存在于细胞膜…中更多。此外,还从膜组分中纯化了未知的mPLC-II。从人血小板膜中分离纯化了两种异三聚体GTP结合蛋白GI2(主成分)和Gi3(Miner成分),并鉴定为百日咳毒素的底物。免疫印迹法检测百日咳毒素不敏感的GTP结合蛋白(GQ)。从人血小板胞浆和细胞膜(c21KG、c25KG、m22KGI、II)中纯化了各种类型的低相对分子质量GTP结合蛋白。经鉴定为smg21A(Krev-1)、Rap1B和Rala。获得了新的C25KG基因,命名为Ram。已知GTP结合蛋白参与凝血酶和血栓素A2介导的人血小板肌醇磷酸的产生。最近有报道血栓素A2受体结合百日咳毒素不敏感GTP结合蛋白(GQ)和PLC-β1被GQα特异性激活。我们先前观察到人血小板膜相关PLC被GI和GO激活。PLC-β2同工酶是GTP结合蛋白的活化β-γ亚基。凝血酶受体与两种不同的GTP结合蛋白(GI和GQ)偶联。提示血栓素A2刺激通过GQ激活PLC-β1,凝血酶诱导GI2β-γ亚基介导的PLC(β2)激活。我们研究了胞浆组份中PLC-γ与肌动蛋白-明胶蛋白复合体的关系,提示明胶蛋白可能在调节人血小板PLC活性中发挥作用。较少
英文摘要
It has generally been known that the stimulation of phosphoinositide turnover after the agonist-receptor interaction is initiated by the activation of phosphoinositide-specific phospholipase C (PI-PLC) through putative GTP-binding protein in plasma membranes. Blood platelets have frequently been used as a potentially useful model for studying transmembrane signaling. The purpose of this study was to clarify the mechanism of activation of thePI- PLC in human platelets, where PI-PLC isozymes and GTP-binding proteins were purified and reconstituted with adequate combination. The effective resolution of PI-PLCs of human platelet cytosolic and membrane fractions revealed five distinct activity peaks. The results of Western blotting analysis with various antibodies against PI-PLC isozymes showed that PLC-beta, PLC-gamma and PLC-delta isozymes were identified and two other unidentified activity peaks were separated from the cytosolic fraction and PLC-beta was mainly conatined in the membranes … More . Furthermore, unidentified mPLC-II was purified from the membrane fractions. Two heterotrimeric GTP-binding proteins, Gi2 (main component) and Gi3 (miner component) were purified and identified as substrates of pertussis toxin from human platelet membranes. The purtussis toxin-insensitive GTP-binding protein (Gq) was detected by Western blotting analysis with anti-GLalpha antibody. Various types of low molecular weight GTP- binding proteins were purified from cytosol and membrane of human platelets (c21KG, c25KG, m22KGI,II). These are identified to be smg21A (Krev-1), rap1B and ralA. The cDNA of novel c25KG was obtained and namedas ram. GTP-binding proteins have been known to be involved in thrombin and thromboxane A2-mediated production of inositol phosphates in human platelets. It has been recently reported that thromboxane A2 receptor bound pertussis toxin-insensitive GTP-binding protein (Gq) and PLC-beta 1 was specifically activated by the Gq alpha . We previously observed that human platelet membrane-associated PLC was activated by Gi and Go. The PLC-beta2 isozyme was activated beta gamma subunit of GTP-binding protein. Thrombin receptor coupled with two different GTP-binding proteins (Gi and Gq). These results suggested that thromboxane A2-stimulation activate PLC-beta1 via Gq and thrombin induced the PLC (beta2) activation mediated by beta gamma subunit of Gi2. We investigated that PLC-gamma was associated with actin-gelsolin complex in cytosolic fraction and suggested that gelsolin may play a role in regulation of PLC activity in human platelets. Less
期刊论文(92)
专著(0)
科研奖励(0)
会议论文
Nakashima,S.: "Genisteine,a protein tyrosin kinase inhibitor,inhibits thromboxane A2ーmediated humna platlet responses" Mol.Pharmacol. 39. 475-480 (1991)
Nakashima, S.:“Genisteine,一种蛋白酪氨酸激酶抑制剂,抑制血栓素 A2 介导的人类血小板反应”Mol.Pharmacol 39. 475-480 (1991)
DOI: --
发表时间:
期刊:
影响因子: --
作者: []
通讯作者:
Banno,Y.: "Isolation of a polyphosphoinositideーphospholipase C(Type)" Platelets.
Banno, Y.:“多磷酸肌醇-磷脂酶 C(类型)的分离”血小板。
DOI: --
发表时间:
期刊:
影响因子: --
作者: []
通讯作者:
Y.Banno: "Effects of gelsoin on human platelet cytosolic phosphoinositide-phospholipase C isozymes" J.Biol.Chem.267. 6488-6494 (1992)
Y.Banno:“凝溶胶对人血小板胞质磷酸肌醇-磷脂酶 C 同工酶的影响”J.Biol.Chem.267。
DOI: --
发表时间:
期刊:
影响因子: --
作者: []
通讯作者:
S.Nakashima: "Thrombin induces a biphasic 1,2-diacylglycerol production in human platelets." Biochem. J.275. 355-361 (1991)
S.Nakashima:“凝血酶诱导人血小板产生双相 1,2-二酰基甘油。”
DOI: --
发表时间:
期刊:
影响因子: --
作者: []
通讯作者:
共 42 条
    REGULATORY MECHANISM BY PHOSPHOLIPASE D IN OXIDANT-STRESS INDUCED SURVIVAL SIGNALING
    CROSS-TALK OF MEMBRANE LIPID SIGNALING IN CELL DEATH AND SURVIVAL
    MECHNISM OF APOPTOSIS INDUCED BY MEMBRANE LIPID SYGNALING
    • 批准号:
      12470042
    • 项目类别:
      Grant-in-Aid for Scientific Research (B)
    • 资助金额:
      $7.94万
    • 财政年份:
      2000
    • 负责人:
      NOZAWA Yoshinori
    • 依托单位:
    Functional analysis of the new signal transduction enzyme PLD by the molecular genetic technique
    海外基金