Tyrosine phosphorylated c-Cbl regulates platelet functional responses mediated by outside-in signaling

被引:25
作者
Buitrago, Lorena
Langdon, Wallace Y. [2 ]
Sanjay, Archana [3 ,4 ]
Kunapuli, Satya P. [1 ,4 ]
机构
[1] Temple Univ, Sch Med, Dept Physiol, Philadelphia, PA 19122 USA
[2] Univ Western Australia, Sch Pathol & Lab Med, Crawley, Australia
[3] Temple Univ, Dept Anat & Cell Biol, Philadelphia, PA 19122 USA
[4] Temple Univ, Sol Sherry Thrombosis Res Ctr, Philadelphia, PA 19122 USA
基金
美国国家卫生研究院;
关键词
CELL-RECEPTOR STIMULATION; BETA CYTOPLASMIC DOMAIN; SRC-KINASE; GLYCOPROTEIN-VI; PHOSPHATIDYLINOSITOL; 3-KINASE; CYTOSKELETAL REORGANIZATION; PHOSPHOINOSITIDE; OSTEOCLAST SURVIVAL; FAMILY PROTEINS; BINDING-SITE;
D O I
10.1182/blood-2011-01-328807
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
c-Cbl protein functions as an E3 ligase and scaffolding protein, where 3 residues, Y700, Y731, and Y774, upon phosphorylation, have been shown to initiate several signaling cascades. In this study, we investigated the role of these phospho-tyrosine residues in the platelet functional responses after integrin engagement. We observed that c-Cbl Y700, Y731 and Y774 undergo phosphorylation upon platelet adhesion to immobilized fibrinogen, which was inhibited in the presence of PP2, a pan-src family kinase (SFK) inhibitor, suggesting that c-Cbl is phosphorylated downstream of SFKs. However, OXSI-2, a Syk inhibitor, significantly reduced c-Cbl phosphorylation at residues Y774 and Y700, without affecting Y731 phosphorylation. Interestingly, PP2 inhibited both platelet-spreading on fibrinogen as well as clot retraction, whereas OXSI-2 blocked only platelet-spreading, suggesting a differential role of these tyrosine residues. The physiologic role of c-Cbl and Y731 was studied using platelets from c-Cbl KO and c-Cbl(YF/YF) knock-in mice. c-Cbl KO and c-Cbl(YF/YF) platelets had a significantly reduced spreading over immobilized fibrinogen. Furthermore, clot retraction with c-Cbl KO and c-Cbl(YF/YF) platelets was drastically delayed. These results indicate that c-Cbl and particularly its phosphorylated residue Y731 plays an important role in platelet outside-in signaling contributing to platelet-spreading and clot retraction. (Blood. 2011;118(20):5631-5640)
引用
收藏
页码:5631 / 5640
页数:10
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