Molecular mechanism of vinculin activation and nanoscale spatial organization in focal adhesions.
Molecular mechanism of vinculin activation and nanoscale spatial organization in focal adhesions.
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DOI：
10.1038/ncb3180
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发表时间：
2015-07
影响因子：
21.3
通讯作者：
Waterman, Clare M.
中科院分区：
文献类型：
--
作者：
Case, Lindsay B.;Baird, Michelle A.;Shtengel, Gleb;Campbell, Sharon L.;Hess, Harald F.;Davidson, Michael W.;Waterman, Clare M.
粘着斑（Focal adhesions，FA）将细胞外基质（extracellular matrix，ECM）连接到肌动蛋白细胞骨架以介导细胞粘附、迁移、机械传感和信号传导。FA具有保守的纳米级蛋白质组织，表明FA内蛋白质的位置调节其活性和功能。黏着斑蛋白结合不同的FA蛋白来介导不同的细胞功能，但黏着斑蛋白的相互作用如何在FA中时空组织尚不清楚。使用干涉光激活定位（iPALM）超分辨率显微镜来测定粘着斑蛋白纳米级定位和FRET生物传感器来测定粘着斑蛋白构象，我们发现，在FA成熟过程中，FA内向上重新定位有利于粘着斑蛋白激活和机械强化FA。非活性黏着斑蛋白通过结合磷酸桩蛋白定位于FA中的较低整合素信号传导层。Talin结合激活黏着斑蛋白并靶向FA中较高的活性黏着斑蛋白，其中黏着斑蛋白可以参与逆行肌动蛋白流动。因此，特定的蛋白质相互作用在纳米尺度上在FA内空间分离，以调节黏着斑蛋白的激活和功能。
Focal adhesions (FAs) link the extracellular matrix (ECM) to the actin cytoskeleton to mediate cell adhesion, migration, mechanosensing and signaling. FAs have conserved nanoscale protein organization, suggesting that the position of proteins within FAs regulates their activity and function. Vinculin binds different FA proteins to mediate distinct cellular functions, but how vinculin’s interactions are spatiotemporally organized within FA is unknown. Using interferometric photo-activation localization (iPALM) super-resolution microscopy to assay vinculin nanoscale localization and a FRET biosensor to assay vinculin conformation, we found that upward repositioning within the FA during FA maturation facilitates vinculin activation and mechanical reinforcement of FA. Inactive vinculin localizes to the lower integrin signaling layer in FA by binding to phospho-paxillin. Talin binding activates vinculin and targets active vinculin higher in FA where vinculin can engage retrograde actin flow. Thus, specific protein interactions are spatially segregated within FA at the nano-scale to regulate vinculin activation and function.
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影响因子：
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作者：
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