Center for Soft Matter Research, Department of Physics, New York University, New York, New York; Molecular Biophysics and Biochemistry Training Program, NYU Grossman School of Medicine, New York, New York.
Center for Soft Matter Research, Department of Physics, New York University, New York, New York.
Biophys J. 2023 Sep 5;122(17):3506-3515. doi: 10.1016/j.bpj.2023.07.026. Epub 2023 Jul 31.
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 both trans (apposed cell) and cis (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-cadherin trans binding, whereas a high protein surface concentration enables the cis-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 lateral cis interaction with a "cis mutant" 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.
E-钙黏蛋白在细胞间黏附中起着核心作用。野生型钙黏蛋白的胞外结构域通过跨(对向细胞)和顺(同一细胞)相互作用在细胞-细胞界面上形成结晶状二维晶格。除了这些细胞外的力,黏附强度还进一步受到涉及α和β连环蛋白介导的钙黏蛋白与肌动蛋白细胞骨架之间相互作用的细胞质现象的调节。细胞-细胞黏附可以通过尚未在分子细节上明确表征的机制在张力下进一步增强。在这里,我们定量确定了钙黏蛋白胞外结构域在机械感应中的作用。为此,我们设计了一种 E-钙黏蛋白包被乳液系统,其中液滴表面张力通过与蛋白质结合强度平衡来产生稳定的黏附区域。为了达到蜂窝/内聚极限,通过离心对初始乳液进行压缩有助于 E-钙黏蛋白的跨结合,而高蛋白质表面浓度能够增强界面的顺式增强稳定性。我们观察到一个突然的浓度依赖性,在具有恒定结晶密度的黏附物中招募,类似于一阶相变。用“顺式突变体”去除侧向顺式相互作用会将这种转变转移到更高的表面密度,导致更密集但更弱的黏附。在这两种蛋白质中,对变形更大区域的稳定与单分子实验一致,该实验表明钙黏蛋白胞外结构域的寿命在力的作用下增强,这可能归因于“X-二聚体”键。