Mechanical regulation of protein structure and function
We are developing technology for directly visualizing the effects of force on the conformation of actin filaments and their interactions with binding partners. We use approaches ranging from optical tweezers to a myosin molecular motor-based reconstitution system compatible with both fluorescence microscopy functional assays and cryo-EM structural studies. In the longer term we are working towards building general platforms for visualizing the effects of forces on macromolecular structure.
Cryo-EM visualizes how bending alters F-actin structure by repositioning subunits in the filament lattice, which is modulated by actin nucleotide state. From Reynolds et al., Nature, 2022.
TIRF movie of the LIM domain protein FHL3 binding in patches along actin filaments in the presence of myosin-generated forces. From Sun et al., Dev. Cell, 2020.
Simultaneous optical trap / confocal fluorescence assay, developed in collaboration with Shixin Liu's lab, demonstrates enhanced binding of α-catenin to single actin filaments in the presence of tension. From Mei et al., Elife, 2020.