Cryo-EM Studies of Dynamin Assembled on Membranes In Vitro and within Cells

Abstract

Dynamin GTPases enable endocytosis by wrapping around the neck of budding vesicles to catalyze membrane fission. While structural and biophysical studies have contributed to elucidating the mechanism of dynamin-mediated membrane fission, questions remain about how dynamin forms the final pre-fission super-constricted state and how dynamin assembles in cells. To address these questions, we have applied cryo-electron microscopy (cryo-EM) helical reconstruction methods, time-resolved cryo-EM, and cryo-electron tomography (cryo-ET) to elucidate the assembly of dynamin in vitro on lipid tubules and in vivo on membranes within HeLa cells.