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Role of ADP ribosylation factor (ARF) in platelet fanctions

ADP 核糖基化因子 (ARF) 在血小板功能中的作用

基本信息

批准号：
08671246
负责人：
TERUI Takeshi
金额：
$ 1.28万
依托单位：
SAPPORO MEDICAL UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1996
资助国家：
日本
起止时间：
1996 至 1997
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08671246/
关键词：
ADP ribosylation factor phospholipase D pletelet brefeldine A (BFA)beta-thromboglobulin aggregation ADP collagen 血小板凝集放出反応 Breferdin A 低分子量G蛋白ホスファチジン酸 PIP2 ホスファチジルコリン

项目摘要

Stimulation of phospholipase D (PLD) after activation of cell surface receptors has been reported in many cell types. We have investigated the mechanisms of activation of this enzyme, especially ADP ribosylation factor (ARF) dependent PLD,by ADP or collagen in human pletelet. aggregation is still unclearinduced with agonists has been studied.We examined the expression of ADP ribosylation factor (ARF) in pletelets by immuboblotting and ARF dependent type PLD (PLD1) by RP-PCR.Then, we revealed that both ADP and collagen indused ARF1 activation ([^<35>S]GTPgS binding) on pletelets membrane upto 10-fold. PLD activity on platelet membrane was stimulated 4- to 5- fold by the addition of GTPgammaS and myrisoylated recombinant ARF1. And both ADP and collagen also induced ARF dependent PLD activity about 150%.Brefeldine A (BFA) inhibits activation of ARF and PLD about 40% and 50% respectively, aggregation of platelets by ADP,however, was not inibited so much (only 25% inhibition). However, BFA inhibited 50% of maxmal aggreation of platelets by collagen. Considering ADP induces primary aggregation and collagen induces secondary aggregation of platelete, BFA might have inhibited granule relese and so on. BFA inhibited 60% of beta-TG release from platelet.In conclution, ARF-PLD signaling seems to be under GPIa/IIa or GPIb/V/IX as collagen receptor and work as a signal transduction for granule release and platelet aggregation.

磷脂酶D（PLD）的刺激后，细胞表面受体的活化已被报道在许多细胞类型。我们研究了ADP或胶原激活人血小板中该酶，特别是ADP核糖基化因子（ARF）依赖的PLD的机制。我们用免疫印迹法检测了ADP核糖基化因子（ADP ribosylation factor，ARF）的表达，用RP-PCR法检测了ARF依赖型PLD（PLD 1）的表达，发现ADP和胶原均可诱导ARF 1活化（[^<35>S]GTPgS结合）达10倍以上。血小板膜上的PLD活性通过加入GTP γ S和豆蔻酰化重组ARF 1刺激4- 5倍。ADP和胶原均可诱导ARF依赖的PLD活性约150%，而BFA可分别抑制ARF和PLD活性约40%和50%，但对ADP诱导的血小板聚集无明显抑制作用（仅抑制25%）。而BFA对胶原诱导的血小板最大聚集有50%的抑制作用。考虑到ADP诱导血小板的初级聚集，胶原诱导血小板的次级聚集，BFA可能具有抑制血小板颗粒释放等作用，BFA抑制60%的β-TG释放，提示ARF-PLD信号通路可能作为胶原受体在GPIa/IIa或GPIb/V/IX的调控下，参与血小板颗粒释放和聚集的信号转导。