权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Regulation mechanism of phagocytotic process in leukocytes by an actin-binding protein, p57, and immune diseases

肌动蛋白结合蛋白p57对白细胞吞噬过程的调节机制与免疫疾病

基本信息

批准号：
10672064
负责人：
TOYOSHIMA Satoshi
金额：
$ 1.41万
依托单位：
Hoshi University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10672064/
关键词：
actin-binding protein, p57 leukocytes phagocytes phagosome acin NADPH osidase Protein kinase C X-linked chronic granulomatous disease 蛋白質リン酸化リソソーム

项目摘要

In the present study, intracellular translocation of the actin-binding protein, p57, in phagocytotic leukocytes and its mechanism were investigated. First, two anitbodies with defferent specificities were prepared for the detection of intracellular p57. Ingestion of opsonized zymosan (OpZ) in neutrophils began within 30 seconds of particle binding and forming phagosomes were enriched for both F-action and p57. However, F-actin and p57 were shed from nascent phagosomes once particle internalization was complete (【approximately equal】5 MINUTES). On the other hand, NADPH oxidase subunits p47phox and p67phox were also recruited to forming phagosomes and were retained on mature phagosomes for at least 15 minutes. These results suggest that p57 plays some role in the formation of phagosomes but not in the expression of mature phagosome function such as superoxide generation. Then, the mechanism of p57 shedding from phagosomes was investigated. Since it has been shown that protein phosphorylation plays a key role in the signaling pathway duraing phagocytosis, phosphorylation of p57 during phagocytosis was studies. Ingestion of OpZ induced a transient increase of p57 phosphorylation. The time of increased phosphorylation corresponded with that of p57 shedding from phagosome. To investigate which protein kinase is responsible for the shedding, effects of inhibitors of protein phosphorylation on it was studied. It was found that protein kinase C inhibitors suppressed the p57 shedding.

本研究探讨了肌动蛋白结合蛋白p57在吞噬性白细胞中的胞内转位及其机制。首先，制备了两种特异性不同的抗体，用于检测细胞内p57。中性粒细胞中调理酵母聚糖（OpZ）的摄入在颗粒结合的30秒内开始，形成的吞噬体富含F-作用和p57。然而，一旦颗粒内化完成，F-肌动蛋白和p57就会从新生的吞噬体中脱落（[约等于]5 μ TES）。另一方面，NADPH氧化酶亚基p47 phox和p67 phox也被募集以形成吞噬体，并在成熟的吞噬体上保留至少15分钟。这些结果表明，p57在吞噬体的形成中起一定的作用，但在成熟的吞噬体功能，如超氧化物的产生的表达中没有作用。然后，研究了p57从吞噬体脱落的机制。由于蛋白质磷酸化在吞噬过程中的信号转导途径中起着关键作用，因此研究了吞噬过程中p57的磷酸化。摄入OpZ诱导p57磷酸化的瞬时增加。磷酸化增加的时间与p57从吞噬体脱落的时间相对应。为了研究哪种蛋白激酶负责脱落，研究了蛋白磷酸化抑制剂对其的影响。发现蛋白激酶C抑制剂抑制p57脱落。