Transient anchorage of cross-linked glycosyl-phosphatidylinositol-anchored proteins depends on cholesterol, Src family kinases, caveolin, and phosphoinositides
Transient anchorage of cross-linked glycosyl-phosphatidylinositol-anchored proteins depends on cholesterol, Src family kinases, caveolin, and phosphoinositides
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DOI：
10.1083/jcb.200512116
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发表时间：
2006-10-09
影响因子：
7.8
通讯作者：
Jacobson, Ken
中科院分区：
文献类型：
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作者：
Chen, Yun;Thelin, William R.;Jacobson, Ken
外莱亚。ET质膜成分，包括糖基磷脂酰肌醇锚定蛋白（GPIAPs），将信号传递到细胞内部是膜生物学中的一个开放问题。通过在抗体缀合的40-nm金颗粒下有意地交联几种GPIAP，金颗粒诱导的Thy-1和CD 73（5'外肽酶）的簇的瞬时锚定在成纤维细胞中发生300 ms至10 s的时间。通过胆固醇消耗、添加Src家族激酶（SFK）抑制剂PP 2或Src-Yes-Fyn敲除细胞消除瞬时锚定。Caveolin-1基因敲除细胞表现出短暂的锚定时间减少，表明部分参与的Caveolin-1。相比之下，跨膜蛋白，囊性纤维化跨膜传导调节，表现出短暂的锚定，发生没有故意增强交联，而且，它只是轻微抑制胆固醇耗竭或SFK抑制，完全依赖于其PDZ结合结构域与细胞骨架适配器EBP 50的相互作用。我们建议，交联GPIAPs成为瞬时锚定通过胆固醇依赖性SFK调节跨膜簇传感器和细胞骨架之间的联系。
ow outer lea. et plasma membrane components, including glycosyl-phosphatidylinositol-anchored proteins (GPIAPs), transmit signals to the cell interior is an open question in membrane biology. By deliberately cross-linking several GPIAPs under antibody-conjugated 40-nm gold particles, transient anchorage of the gold particle - induced clusters of both Thy-1 and CD73, a 5' exonucleotidase, occurred for periods ranging from 300 ms to 10 s in fibroblasts. Transient anchorage was abolished by cholesterol depletion, addition of the Src family kinase (SFK) inhibitor PP2, or in Src-Yes-Fyn knockout cells. Caveolin-1 knockout cells exhibited a reduced transient anchorage time, suggesting the partial participation of caveolin-1. In contrast, a transmembrane protein, the cystic fibrosis transmembrane conductance regulator, exhibited transient anchorage that occurred without deliberately enhanced cross-linking; moreover, it was only slightly inhibited by cholesterol depletion or SFK inhibition and depended completely on the interaction of its PDZ-binding domain with the cytoskeletal adaptor EBP50. We propose that cross-linked GPIAPs become transiently anchored via a cholesterol-dependent SFK-regulatable linkage between a transmembrane cluster sensor and the cytoskeleton.