Push-pull mechanics of E-cadherin ectodomains in biomimetic adhesions
Push-pull mechanics of E-cadherin ectodomains in biomimetic adhesions
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E-钙粘蛋白胞外域在仿生粘连中的推拉力学
DOI：
10.1016/j.bpj.2023.07.026
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发表时间：
2023
影响因子：
3.4
通讯作者：
Brujic, Jasna
中科院分区：
文献类型：
--
作者：
Nagendra, Kartikeya;Izzet, Adrien;Judd, Nicolas B.;Zakine, Ruben;Friedman, Leah;Harrison, Oliver J.;Pontani, Léa-Laetitia;Shapiro, Lawrence;Honig, Barry;Brujic, Jasna
E-钙粘蛋白在细胞-细胞粘附中起核心作用。野生型钙粘蛋白的胞外域在细胞-细胞界面上形成一个晶体样的二维晶格，这种晶格由反式（并列细胞）和顺式（相同细胞）相互作用介导。除了这些细胞外的力量，粘附强度进一步调节胞质现象，涉及α和β连环蛋白介导的钙粘蛋白和肌动蛋白细胞骨架之间的相互作用。细胞-细胞粘附在张力下可以通过尚未在分子细节上明确表征的机制进一步加强。在这里，我们定量确定的钙粘蛋白胞外域在机械传感的作用。为此，我们设计了一种E-钙粘蛋白包被的乳液系统，其中液滴表面张力通过蛋白质结合强度来平衡，以产生稳定的粘附区域。为了达到蜂窝/内聚极限，通过离心进行的初始乳液压缩促进E-钙粘蛋白transbinding，而高蛋白质表面浓度使界面的thecis增强的稳定性成为可能。我们观察到浓度对招募到恒定晶体密度的粘连的突然依赖性，让人想起一级相变。去除与“顺式突变体”的横向顺式相互作用，将这种过渡转移到更高的表面密度，导致更致密，但更弱的粘附。在这两种蛋白质中，逐渐变大的变形区域的稳定性与单分子实验一致，该实验显示钙粘蛋白胞外域中的力依赖性寿命增强，这可能归因于“X-二聚体”键。
E-cadherin plays a central role in cell-cell adhesion. The ectodomains of wild-type cadherins form a crystalline-like two-dimensional lattice in cell-cell interfaces mediated by bothtrans(apposed cell) andcis(same cell) interactions. In addition to these extracellular forces, adhesive strength is further regulated by cytosolic phenomena involving α and β catenin-mediated interactions between cadherin and the actin cytoskeleton. Cell-cell adhesion can be further strengthened under tension through mechanisms that have not been definitively characterized in molecular detail. Here we quantitatively determine the role of the cadherin ectodomain in mechanosensing. To this end, we devise an E-cadherin-coated emulsion system, in which droplet surface tension is balanced by protein binding strength to give rise to stable areas of adhesion. To reach the honeycomb/cohesive limit, an initial emulsion compression by centrifugation facilitates E-cadherintransbinding, whereas a high protein surface concentration enables thecis-enhanced stabilization of the interface. We observe an abrupt concentration dependence on recruitment into adhesions of constant crystalline density, reminiscent of a first-order phase transition. Removing the lateralcisinteraction with a "cismutant" shifts this transition to higher surface densities leading to denser, yet weaker adhesions. In both proteins, the stabilization of progressively larger areas of deformation is consistent with single-molecule experiments that show a force-dependent lifetime enhancement in the cadherin ectodomain, which may be attributed to the "X-dimer" bond.