Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama University , Okayama 700-0082, Japan.
Stanford Cardiovascular Institute, Stanford University , 265 Campus Drive, Stanford, California 94305, United States.
ACS Nano. 2016 Dec 27;10(12):10745-10752. doi: 10.1021/acsnano.6b03314. Epub 2016 Nov 28.
Integrins mediate cell adhesion to the extracellular matrix and enable the construction of complex, multicellular organisms, yet fundamental aspects of integrin-based adhesion remain poorly understood. Notably, the magnitude of the mechanical load experienced by individual integrins within living cells is unclear, due principally to limitations inherent to existing techniques. Here we use Förster resonance energy transfer-based molecular tension sensors to directly measure the distribution of loads experienced by individual integrins in living cells. We find that a large fraction of integrins bear modest loads of 1-3 pN, while subpopulations bearing higher loads are enriched within adhesions. Further, our data indicate that integrin engagement with the fibronectin synergy site, a secondary binding site specifically for αβ integrin, leads to increased levels of αβ integrin recruitment to adhesions but not to an increase in overall cellular traction generation. The presence of the synergy site does, however, increase cells' resistance to detachment by externally applied loads. We suggest that a substantial population of integrins experiencing loads well below their peak capacities can provide cells and tissues with mechanical integrity in the presence of widely varying mechanical loads.
整合素介导细胞与细胞外基质的黏附,并使复杂的多细胞生物得以构建,但整合素黏附的基本方面仍知之甚少。值得注意的是,由于现有技术固有的限制,单个整合素在活细胞中所经历的机械负载的大小尚不清楚。在这里,我们使用基于Förster 共振能量转移的分子张力传感器来直接测量活细胞中单个整合素所经历的负载分布。我们发现,很大一部分整合素承载着适度的 1-3 pN 的负载,而承载更高负载的亚群在黏附处富集。此外,我们的数据表明,整合素与纤维连接蛋白协同结合位点(专门针对 αβ 整合素的二级结合位点)的结合会导致 αβ 整合素募集到黏附处的水平增加,但不会导致整体细胞牵引力的增加。然而,协同结合位点的存在确实会增加细胞对外部施加负载的抵抗能力。我们认为,在存在广泛变化的机械负载的情况下,大量经历负载远低于其峰值容量的整合素可以为细胞和组织提供机械完整性。
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