Flotillins interact with PSGL-1 in neutrophils and, upon stimulation, rapidly organize into membrane domains subsequently accumulating in the uropod.
Flotillins interact with PSGL-1 in neutrophils and, upon stimulation, rapidly organize into membrane domains subsequently accumulating in the uropod.
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DOI：
10.1371/journal.pone.0005403
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发表时间：
2009
期刊：
影响因子：
3.7
通讯作者：
Niggli V
中科院分区：
文献类型：
--
作者：
Rossy J;Schlicht D;Engelhardt B;Niggli V
中性粒细胞响应趋化因子而增殖和迁移。不同类型的膜微区（筏）已被假定存在于后方和前方的极化白细胞和破坏筏胆固醇隔离防止白细胞极化。Reggie/flotillin-1和-2是两种高度同源的蛋白质，其普遍富集在耐洗涤剂膜中，并且被认为通过形成同源寡聚体和异源寡聚体来形成膜微区。本研究的目的是研究中性粒细胞活化过程中动态膜微区重组。我们现在显示，使用免疫荧光染色和免疫共沉淀，内源性flotillin-1和-2共定位和关联在静止的球形和极化的初级中性粒细胞。在化学引诱物刺激后，漂浮蛋白很早就重新分布，并在90%以上的中性粒细胞中形成明显的帽。在稍后的时间点，漂浮蛋白积聚在极化细胞的尾足中。趋化肽诱导的再分布和封端的flotillin需要完整性和动态的肌动蛋白细胞骨架，但不涉及Rho激酶依赖的信号相关的尾足的形成。两种flotillin亚型都参与了该膜结构域的形成，因为与仅表达一种标记亚型的细胞相比，在分化的HL-60细胞中标记flotillin-1和-2的共过表达显著增强了外源表达的flotillins的尾足定位。Flotillin-1和-2与P-选择素糖蛋白配体1（PSGL-1）在静息和刺激的中性粒细胞，如共定位和免疫共沉淀所示。从PSGL-1缺陷型小鼠分离的中性粒细胞表现出与从野生型动物分离的细胞相同程度的flotillin帽，这意味着PSGL-1不是形成flotillin帽所必需的。最后，我们表明，刺激依赖的重新分配的其他尾足定位的蛋白质，CD 43和埃兹蛋白/radixin/膜突蛋白，发生慢得多的flotillins和PSGL-1。这些结果表明，富浮体蛋白肌动蛋白依赖性膜微区的重要参与中性粒细胞尾足的形成和/或稳定和组织PSGL-1的尾足定位。
Neutrophils polarize and migrate in response to chemokines. Different types of membrane microdomains (rafts) have been postulated to be present in rear and front of polarized leukocytes and disruption of rafts by cholesterol sequestration prevents leukocyte polarization. Reggie/flotillin-1 and -2 are two highly homologous proteins that are ubiquitously enriched in detergent resistant membranes and are thought to shape membrane microdomains by forming homo- and hetero-oligomers. It was the goal of this study to investigate dynamic membrane microdomain reorganization during neutrophil activation. We show now, using immunofluorescence staining and co-immunoprecipitation, that endogenous flotillin-1 and -2 colocalize and associate in resting spherical and polarized primary neutrophils. Flotillins redistribute very early after chemoattractant stimulation, and form distinct caps in more than 90% of the neutrophils. At later time points flotillins accumulate in the uropod of polarized cells. Chemotactic peptide-induced redistribution and capping of flotillins requires integrity and dynamics of the actin cytoskeleton, but does not involve Rho-kinase dependent signaling related to formation of the uropod. Both flotillin isoforms are involved in the formation of this membrane domain, as uropod location of exogenously expressed flotillins is dramatically enhanced by co-overexpression of tagged flotillin-1 and -2 in differentiated HL-60 cells as compared to cells expressing only one tagged isoform. Flotillin-1 and -2 associate with P-selectin glycoprotein ligand 1 (PSGL-1) in resting and in stimulated neutrophils as shown by colocalization and co-immunoprecipitation. Neutrophils isolated from PSGL-1-deficient mice exhibit flotillin caps to the same extent as cells isolated from wild type animals, implying that PSGL-1 is not required for the formation of the flotillin caps. Finally we show that stimulus-dependent redistribution of other uropod-located proteins, CD43 and ezrin/radixin/moesin, occurs much slower than that of flotillins and PSGL-1. These results suggest that flotillin-rich actin-dependent membrane microdomains are importantly involved in neutrophil uropod formation and/or stabilization and organize uropod localization of PSGL-1.
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筏结构域的动态重新分布是细胞趋化过程中信号传导的组织平台。
影响因子：
7.8
作者：
Gómez-Moutón, C;Lacalle, RA;Mira, E;Jiménez-Baranda, S;Barber, DF;Carrera, AC;Martínez, C;Mañes, S
通讯作者：
Mañes, S
影响因子：
3.5
作者：
Langhorst, Matthias F.;Solis, Gonzalo P.;Stuermer, Claudia A. O.
通讯作者：
Stuermer, Claudia A. O.
影响因子：
4.8
作者：
Langhorst, Matthias F.;Reuter, Alexander;Stuermer, Claudia A. O.
通讯作者：
Stuermer, Claudia A. O.
影响因子：
15.9
作者：
LORANT, DE;MCEVER, RP;ZIMMERMAN, GA
通讯作者：
ZIMMERMAN, GA
影响因子：
16.8
作者：
Mañes, S;Lacalle, RA;Martínez, C
通讯作者：
Martínez, C