Spatial distribution and functional significance of activated vinculin in living cells.
Spatial distribution and functional significance of activated vinculin in living cells.
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DOI：
10.1083/jcb.200410100
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发表时间：
2005-05-09
期刊：
影响因子：
--
通讯作者：
Craig SW
中科院分区：
文献类型：
--
作者：
Chen H;Cohen DM;Choudhury DM;Kioka N;Craig SW
构象变化被认为是重要的粘着斑蛋白的功能，在网站的细胞粘附。然而，对黏着斑蛋白在活细胞中的构象一无所知。使用福斯特共振能量转移探针，报告纽蛋白的构象的变化，我们发现，纽蛋白是在肌动蛋白结合的构象在一个外围带的粘附点在扩展细胞。然而，在具有既定极性的完全展开的细胞中，黏着斑蛋白的构象在粘着斑处是可变的。延时成像揭示了收缩区黏着斑蛋白丢失之前的构象变化梯度。在稳定或突出的区域，黏着斑蛋白的募集不一定与肌动蛋白结合构象偶联。然而，黏着斑蛋白构象的不同测量，活化黏着斑蛋白对vinexin β的募集，表明内源性黏着斑蛋白的自身抑制在局灶性粘连处放松。除了提供黏着斑蛋白在局部粘连处被激活的直接证据外，这项研究还表明特定的功能构象与区域细胞动力学相关。
Conformational change is believed to be important to vinculin's function at sites of cell adhesion. However, nothing is known about vinculin's conformation in living cells. Using a Forster resonance energy transfer probe that reports on changes in vinculin's conformation, we find that vinculin is in the actin-binding conformation in a peripheral band of adhesive puncta in spreading cells. However, in fully spread cells with established polarity, vinculin's conformation is variable at focal adhesions. Time-lapse imaging reveals a gradient of conformational change that precedes loss of vinculin from focal adhesions in retracting regions. At stable or protruding regions, recruitment of vinculin is not necessarily coupled to the actin-binding conformation. However, a different measure of vinculin conformation, the recruitment of vinexin β by activated vinculin, shows that autoinhibition of endogenous vinculin is relaxed at focal adhesions. Beyond providing direct evidence that vinculin is activated at focal adhesions, this study shows that the specific functional conformation correlates with regional cellular dynamics.
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影响因子：
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