PAF-STIMULATED PHOSPHOINOSITIDE TURNOVER IN PLATELETS
PAF 刺激血小板中的磷酸肌醇周转
基本信息
- 批准号:3072519
- 负责人:
- 金额:$ 5.4万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:1989
- 资助国家:美国
- 起止时间:1989-09-01 至 1994-08-31
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
The candidate has developed an everlasting interest in membranes,
phosphoinositides (PPI) and phospholipases. The immediate goal is to
build a foundation for the research project outlined in this application
which will enable, in the long term, to unravel the importance of PPI
and phospholipases in cellular functions.
Project: Breakdown of PPI by phospholipase C (PLC) in various cells,
including platelets, is involved in signal transduction. The HYPOTHESIS
is that (a) platelet activating factor (PAF) receptor is coupled via a
G-protein to the activation and regulation of the PLC in platelets and
(b)exogenous PLC or activation of endogenous PLC cause release of PI-
anchored proteins from platelet membrane surfaces. There are four
experimental plans.
[I] TO DETERMINE MECHANISM(S) OF PAF RECEPTOR-COUPLED ACTIVATION AND
DESENSITIZATION OF PLC: (A) to correlate [3H]PAF binding to PLC
activation in desensitized platelets as compared to control; (B) to
manipulate G-proteins by using NaF, GTPgammas and pertussis toxin and to
correlate their influence on PLC activation; (C) to correlate protein
kinase C-mediated phosphorylation to the activation and desensitization
of PLC.
[II] TO DETERMINE MOLECULAR INTERACTIONS AMONG PLC, G-PROTEINS AND PAF
RECEPTOR: Platelet membranes will be solubilized and fractionated by
column chromatography. [3H]PAF binding, GTPase, 32P-GTP binding and PLC
activities will be monitored in fractions to establish their functional
associations.
[III] TO DETERMINE IMPORTANCE OF PAF RECEPTOR COUPLED ACTIVATION OF PLC
IN THE HYPERSENSITIVITY OF DIABETIC HUMAN PLATELETS.
[IV] TO DETERMINE THE RELEASE OF PI-ANCHORED MEMBRANE
PROTEIN/GLYCOPROTEIN BY EXOGENOUS AND ENDOGENOUS PLC: Release of
proteins by PLC (B. thuringiensis) from labelled platelets;
identification by SDS-PAGE/fluirography and antibodies; use of
differential phase partitioning to monitor PI-anchored proteins after
PAF stimulation.
The project will provide novel insight(s) in PAF-stimulated PPI turnover
and on the importance of PI-anchored proteins in platelets.
这位候选人对膜产生了持久的兴趣,
磷脂酶(PPI)和磷脂酶。眼下的目标是
为本申请中概述的研究项目奠定基础
从长远来看,这将使PPI的重要性得到揭示
以及细胞功能中的磷脂酶。
项目:磷脂酶C(PLC)在不同细胞中分解PPI,
包括血小板在内,都参与信号转导。假说
是(A)血小板活化因子(PAF)受体通过
G蛋白对血小板内PLC的激活和调节
(B)外源性PLC或内源性PLC的激活导致PI的释放-
从血小板膜表面锚定的蛋白质。一共有四个
实验计划。
[I]确定PAF受体偶联激活的机制(S)
PLC的脱敏作用:(A)[~3H]PAF与PLC结合的相关性
与对照组相比,脱敏血小板的激活;
利用NaF、GTP和百日咳毒素操纵G蛋白,并
关联它们对PLC激活的影响;(C)关联蛋白质
激酶C介导的磷酸化对激活和脱敏的影响
可编程逻辑控制器。
[II]确定PLC、G-蛋白和PAF之间的分子相互作用
受体:血小板膜将被溶解和分级
柱层析法。PAF结合、GTP酶、32P-GTP结合和PLC
将对部分活动进行监控,以确定其功能
联想。
[III]确定PAF受体偶联激活PLC的重要性
糖尿病人血小板的超敏反应。
[IV]测定PI锚定膜的释放度
外源性和内源性PLC的蛋白质/糖蛋白:释放
PLC(苏云金芽孢杆菌)从标记的血小板中提取蛋白质;
用十二烷基硫酸钠-PAGE/荧光照相法和抗体鉴定
差示相分割技术监测PI锚定蛋白的研究
PAF刺激。
该项目将为PAF刺激的PPI成交额提供新的见解(S)
以及PI锚定蛋白在血小板中的重要性。
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
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Shivendra D Shukla其他文献
Shivendra D Shukla的其他文献
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{{ truncateString('Shivendra D Shukla', 18)}}的其他基金
Histone Modifications by Ethanol: Mechanism & Consequence
乙醇对组蛋白的修饰:机制
- 批准号:
7424057 - 财政年份:2007
- 资助金额:
$ 5.4万 - 项目类别:
Histone Modifications by Ethanol: Mechanism & Consequence
乙醇对组蛋白的修饰:机制
- 批准号:
7813978 - 财政年份:2007
- 资助金额:
$ 5.4万 - 项目类别:
Histone Modifications by Ethanol: Mechanism & Consequence
乙醇对组蛋白的修饰:机制
- 批准号:
8066791 - 财政年份:2007
- 资助金额:
$ 5.4万 - 项目类别:
Histone Modifications by Ethanol: Mechanism & Consequence
乙醇对组蛋白的修饰:机制
- 批准号:
7266600 - 财政年份:2007
- 资助金额:
$ 5.4万 - 项目类别:
Histone Modifications by Ethanol: Mechanism & Consequence
乙醇对组蛋白的修饰:机制
- 批准号:
7619302 - 财政年份:2007
- 资助金额:
$ 5.4万 - 项目类别:
Histone acetylation by ethanol:Mechanisms & consequence
乙醇乙酰化组蛋白:机制
- 批准号:
6911646 - 财政年份:2004
- 资助金额:
$ 5.4万 - 项目类别:
Histone acetylation by ethanol:Mechanisms & consequence
乙醇乙酰化组蛋白:机制
- 批准号:
6754225 - 财政年份:2004
- 资助金额:
$ 5.4万 - 项目类别:
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