Proximity proteomics identifies septins and PAK2 as decisive regulators of actomyosin-mediated expulsion of von Willebrand factor.
Proximity proteomics identifies septins and PAK2 as decisive regulators of actomyosin-mediated expulsion of von Willebrand factor.
复制标题
DOI：
10.1182/blood.2022017419
复制
发表时间：
2023-02-23
期刊：
影响因子：
20.3
通讯作者：
Nightingale, Thomas D.
中科院分区：
文献类型：
--
作者：
El-Mansi, Sammy;Robinson, Christopher L.;Kostelnik, Katja B.;McCormack, Jessica J.;Mitchell, Tom P.;Lobato-Marquez, Damian;Rajeeve, Vinothini;Cutillas, Pedro;Cutler, Daniel F.;Mostowy, Serge;Nightingale, Thomas D.
邻近蛋白质组学和内皮细胞中的siRNA子筛选揭示了与VWF受调控的肌球蛋白排出相关的蛋白质。PAK2和Septin杂低聚物是肌动球蛋白环功能和有效释放VWF所必需的。作为对组织损伤的反应，在几秒钟内，超大糖蛋白von Willebrand因子(VWF)从内皮存储细胞器(weibel−Palade Body)释放到血管系统的管腔中，在那里它导致血小板的募集。VWF多聚体的显著大小对内皮细胞(ECs)的分泌机制构成了前所未有的负担。ECS已经进化出了克服这一点的机制，最著名的是肌动球蛋白环，它形成、收缩并挤出其笨重的货物。抑制这些结构的形成或功能代表了血栓病理学的一个新的治疗靶点，尽管表征与这种动态过程相关的蛋白质一直是具有挑战性的。我们将APEX2邻近标记与创新的双重功能丧失筛查相结合，以识别与肌动球蛋白环功能相关的蛋白质。我们发现，p21激活的激酶2(PAK2)招募了Septin异源低聚物，这是一种围绕胞外部位形成环的分子相互作用。这一系列事件控制着肌动球蛋白环的功能，有助于有效的胞外释放。PAK2或Septins的遗传或药物抑制导致VWF的低效释放和未能形成血小板捕捉绳。这一新的分子机制为血栓性疾病的控制提供了额外的治疗靶点，并与使用胞外肌动球蛋白机制的其他分泌系统高度相关。Webel-Palade小体(WPB)是内皮细胞特有的分泌细胞器，储存止血蛋白von Willebrand因子以及许多炎症和血管生成介质；调节这些物质的释放对正常止血至关重要。El-Mansi等人通过高通量双功能丧失筛查发现了WPB胞吐和货物排出的新调节器。他们的数据表明，p21激活的激酶2(PAK2)介导的细胞骨架Septin的募集是WPB胞吐的关键步骤，进一步加深了我们对这一重要止血过程的理解。
Proximity proteomics and siRNA subscreens in endothelial cells reveal proteins associated with regulated actomyosin expulsion of VWF. PAK2 and septin hetero-oligomeric complexes are required for actomyosin ring function and effective VWF release. In response to tissue injury, within seconds the ultra-large glycoprotein von Willebrand factor (VWF) is released from endothelial storage organelles (Weibel−Palade bodies) into the lumen of the blood vasculature, where it leads to the recruitment of platelets. The marked size of VWF multimers represents an unprecedented burden on the secretory machinery of endothelial cells (ECs). ECs have evolved mechanisms to overcome this, most notably an actomyosin ring that forms, contracts, and squeezes out its unwieldy cargo. Inhibiting the formation or function of these structures represents a novel therapeutic target for thrombotic pathologies, although characterizing proteins associated with such a dynamic process has been challenging. We have combined APEX2 proximity labeling with an innovative dual loss-of-function screen to identify proteins associated with actomyosin ring function. We show that p21 activated kinase 2 (PAK2) recruits septin hetero-oligomers, a molecular interaction that forms a ring around exocytic sites. This cascade of events controls actomyosin ring function, aiding efficient exocytic release. Genetic or pharmacological inhibition of PAK2 or septins led to inefficient release of VWF and a failure to form platelet-catching strings. This new molecular mechanism offers additional therapeutic targets for the control of thrombotic disease and is highly relevant to other secretory systems that employ exocytic actomyosin machinery. Weibel-Palade bodies (WPB) are endothelial cell–specific secretory organelles that store the hemostatic protein von Willebrand factor, along with many inflammatory and angiogenic mediators; the regulation of release of this cargo is essential for normal hemostasis. El-Mansi et al uncovered new regulators of WPB exocytosis and cargo expulsion through a high-throughput dual loss-of-function screen. Their data identified p21-activated kinase 2 (PAK2)–mediated recruitment of cytoskeletal septin as a crucial step in WPB exocytosis, furthering our understanding of this important hemostatic process.
登录
查看更多内容
影响因子：
16.2
作者：
Hsu, SC;Hazuka, CD;Scheller, RH
通讯作者：
Scheller, RH
DOI：
10.1016/j.biocel.2020.105900
发表时间：
2021-02-01
影响因子：
4
作者：
El-Mansi, Sammy;Nightingale, Thomas D.
通讯作者：
Nightingale, Thomas D.
影响因子：
7.8
作者：
Garcia, Galo, III;Bertin, Aurelie;Nogales, Eva
通讯作者：
Nogales, Eva
影响因子：
56.9
作者：
Dagdas, Yasin F.;Yoshino, Kae;Talbot, Nicholas J.
通讯作者：
Talbot, Nicholas J.
DOI：
10.1161/atvbaha.112.301165
发表时间：
2013-07
期刊：
Arteriosclerosis, thrombosis, and vascular biology
影响因子：
--
作者：
Aslan JE;Itakura A;Haley KM;Tormoen GW;Loren CP;Baker SM;Pang J;Chernoff J;McCarty OJ
通讯作者：
McCarty OJ