Regulation of Traction Force through the Direct Binding of Basigin and Calpain 4.

Traction force and mechanosensing (the ability to sense mechanical attributes of the environment) are two important factors used by a cell to modify behavior during migration. Previously it was determined that the calpain small subunit, calpain 4, regulates the production of traction force independent of its proteolytic holoenzyme. A proteolytic enzyme is formed by calpain4 binding to either of its catalytic partners, calpain 1 and 2. To further understand how calpain 4 regulates traction force, we used two-hybrid analysis to identify more components of the traction pathway. We discovered that basigin, an integral membrane protein and a documented matrix-metalloprotease (MMP) inducer binds to calpain 4 in two-hybrid and pull-down assays. Traction force was deficient when basigin was silenced in MEF cells, and defective in substrate adhesion strength. Consistent with Capn4 MEF cells, the cells deficient in basigin responded to localized stimuli. Together these results implicate basigin in the pathway in which calpain 4 regulates traction force independent of the catalytic large subunits.