WAVE/scars in platelets
WAVE/scars in platelets
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DOI：
10.1182/blood-2003-04-1319
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发表时间：
2005-04-15
期刊：
影响因子：
20.3
通讯作者：
Takenawa, T
中科院分区：
文献类型：
--
作者：
Oda, A;Miki, H;Takenawa, T
使用针对WAVE （WASP [Wiskott-Aldrich综合征蛋白]家族verprolin同源蛋白，也称为Scar）同种异构体的特异性抗体，我们证明人类血小板表达所有3种同种异构体。使用体外下拉技术，胰岛素受体底物p53 （IRSp53）的src同源3 （SH3）结构域比profilin 1更有效地从血小板裂解物中沉淀WAVE2。WAVE1的情况正好相反，两者都没有沉淀WAVE3，这表明WAVE同型体对这些配体具有不同的亲和力，而abl的SH3结构域与所有3种同型体结合。在凝血酶受体激活肽诱导血小板聚集后，3种WAVE亚型分布在富含肌动蛋白的Triton x -100不溶性微球中。我们还发现所有3种WAVE亚型在体内和体外都是钙蛋白酶的底物。虽然这3种同工异构体的一部分通常分布在扩张血小板板足富含肌动蛋白和肌动蛋白相关蛋白2和3 （Arp2/3）的边缘，但在洗涤液提取或细胞固定后，只有WAVE2留在细胞边缘。最后，通过质谱分析，我们发现，据报道与WAVE/Scars相互作用的蛋白质存在于血小板中。这些数据表明，3种WAVE亚型表现出共同而独特的特征，并可能参与血小板中肌动蛋白细胞骨架的调节。(c) 2005年由美国血液学会出版。
Using specific antibodies against isoforms of WAVE (WASP [Wiskott-Aldrich syndrome protein] family Verprolin-homologous protein, also called Scar), we demonstrated that human platelets express all 3 isoforms. With the use of an in vitro pull-down technique, the src homology 3 (SH3) domain of insulin receptor substrate p53 (IRSp53) precipitated WAVE2 from platelet lysates more efficiently than did profilin 1. The opposite was true for WAVE1, and neither precipitated WAVE3, suggesting that WAVE isoforms have different affinities to these ligands, while the SH3 domain of abl binds to all 3 isoforms. The 3 WAVE isoforms were distributed in the actin-rich Triton X-100-insoluble pellets following platelet aggregation induced by thrombin receptor-activating peptide. We also found that all 3 WAVE isoforms are substrates for calpain in vivo and in vitro. Although portions of these 3 isoforms were commonly distributed in the actin-and actin-related protein 2 and 3 (Arp2/3)-rich edge of the lamellipodia in spreading platelets, only WAVE2 remained in the cell fringe following detergent extraction or fixation of the cells. Finally, by mass spectrometry, we found that the proteins, which reportedly interact with WAVE/Scars, are present in platelets. These data suggest that the 3 WAVE isoforms exhibit common and distinct features and may potentially be involved in the regulation of actin cytoskeleton in platelets. (c) 2005 by The American Society of Hematology.