Proximity Mapping of Desmosomes Reveals a Striking Shift in Their Molecular Neighborhood Associated With Maturation.

Desmosomes are multiprotein adhesion complexes that link intermediate filaments to the plasma membrane, ensuring the mechanical integrity of cells across tissues, but how they participate in the wider signaling network to exert their full function is unclear. To investigate this, we carried out protein proximity mapping using biotinylation (BioID). The combined interactomes of the essential desmosomal proteins desmocollin 2a, plakoglobin, and plakophilin 2a (Pkp2a) in Madin-Darby canine kidney epithelial cells were mapped and their differences and commonalities characterized as desmosome matured from Ca2+ dependence to the mature, Ca2+-independent, hyper-adhesive state, which predominates in tissues. Results suggest that individual desmosomal proteins have distinct roles in connecting to cellular signaling pathways and that these roles alter substantially when cells change their adhesion state. The data provide further support for a dualistic concept of desmosomes in which the properties of Pkp2a differ from those of the other, more stable proteins. This body of data provides an invaluable resource for the analysis of desmosome function.

Modulation of endothelial cell migration via manipulation of adhesion site growth using nanopatterned surfaces.

Orthogonally functionalized nanopatterend surfaces presenting discrete domains of fibronectin ranging from 92 to 405 nm were implemented to investigate the influence of limiting adhesion site growth on cell migration. We demonstrate that limiting adhesion site growth to small, immature adhesions using sub-100 nm patterns induced cells to form a significantly increased number of smaller, more densely packed adhesions that displayed few interactions with actin stress fibers. Human umbilical vein endothelial cells exhibiting these traits displayed highly dynamic fluctuations in spreading and a 4.8-fold increase in speed compared to cells on nonpatterned controls. As adhesions were allowed to mature in size in cells cultured on larger nanopatterns, 222 to 405 nm, the dynamic fluctuations in spread area and migration began to slow, yet cells still displayed a 2.1-fold increase in speed compared to controls. As all restrictions on adhesion site growth were lifted using nonpatterned controls, cells formed significantly fewer, less densely packed, larger, mature adhesions that acted as terminating sites for actin stress fibers and significantly slower migration. The results revealed an exponential decay in cell speed with increased adhesion site size, indicating that preventing the formation of large mature adhesions may disrupt cell stability thereby inducing highly migratory behavior.