Inside-out regulation of E-cadherin conformation and adhesion.

Cadherin cell-cell adhesion proteins play key roles in tissue morphogenesis and wound healing. Cadherin ectodomains bind in two conformations, X-dimers and strand-swap dimers, with different adhesive properties. However, the mechanisms by which cells regulate ectodomain conformation are unknown. Cadherin intracellular regions associate with several actin-binding proteins including vinculin, which are believed to tune cell-cell adhesion by remodeling the actin cytoskeleton. Here, we show at the single-molecule level, that vinculin association with the cadherin cytoplasmic region allosterically converts weak X-dimers into strong strand-swap dimers and that this process is mediated by myosin II-dependent changes in cytoskeletal tension. We also show that in epithelial cells, ∼70% of apical cadherins exist as strand-swap dimers while the remaining form X-dimers, providing two cadherin pools with different adhesive properties. Our results demonstrate the inside-out regulation of cadherin conformation and establish a mechanistic role for vinculin in this process.

Biophysical basis of cadherin mediated cell-cell adhesion.

Classical cadherin transmembrane cell-cell adhesion proteins play essential roles in tissue morphogenesis and in mediating tissue integrity. Cadherin ectodomains from opposing cells interact to form load-bearing trans dimers that mechanically couple cells. Cell-cell adhesion is believed to be strengthened by cis clustering of cadherins on the same cell surface. This review summarizes biophysical studies of the structure, interaction kinetics and biomechanics of classical cadherin ectodomains. We first discuss the structure and equilibrium binding kinetics of classical cadherin trans and cis dimers. We then discuss how mechanical stimuli alters the kinetics of cadherin interaction and tunes adhesion. Finally, we highlight open questions on the role of mechanical forces in influencing cadherin structure, function and organization on the cell surface.

Characterizing the Biophysical Properties of Adhesive Proteins in Live Cells Using Single-Molecule Atomic Force Microscopy.

Cell adhesion proteins play essential roles in the formation, regeneration, and maintenance of tissue. However, the molecular mechanisms by which cells regulate the conformation and binding properties of adhesion proteins are poorly understood. These biophysical properties can be resolved, with single-molecule resolution, using atomic force microscopy (AFM). Here, we outline how AFM force measurements can be used to study the conformation, cytoskeletal linkage, binding strength, and force-dependent bond lifetimes of adhesion proteins in live cells.

Cadherins can dimerize via asymmetric interactions.

Cadherins are essential cell-cell adhesion proteins that interact in two distinct conformations: X-dimers and strand-swap dimers. Both X-dimers and strand-swap dimers are thought to exclusively rely on symmetric sets of interactions between key amino acids on both cadherin binding partners. Here, we use single-molecule atomic force microscopy and computer simulations to show that symmetry in cadherin binding is dispensable and that cadherins can also interact in a novel conformation that asymmetrically incorporates key elements of both strand-swap dimers and X-dimers. Our results clarify the biophysical rules for cadherin binding and demonstrate that cadherins interact in a more diverse range of conformations than previously understood.

Single-molecule studies of classical and desmosomal cadherin adhesion.

Classical cadherin and desmosomal cadherin cell-cell adhesion proteins play essential roles in tissue morphogenesis and in maintaining tissue integrity. Deficiencies in cadherin adhesion are hallmarks of diseases like cancers, skin diseases and cardiomyopathies. Structural studies and single molecule biophysical measurements have revealed critical similarities and surprising differences between these key adhesion proteins. This review summarizes our current understanding of the biophysics of classical and desmosomal cadherin adhesion and the molecular basis for their cross-talk. We focus on recent single molecule measurements, highlight key insights into the adhesion of cadherin extracellular regions and their relation to associated diseases, and identify major open questions in this exciting area of research.