Fig. 1: Purification and characterization of MBP-Dyn1ΔPRD.

a Schematic of dynamin 1 constructs involved in representative structural studies (PDB codes on the left). The PRD domain is unstructured, removed for most structural studies. 7AX3 is the construct used in this study, showing a two-start helix in complex with GMP-PCP in the absence of lipid. 6DLU is the one-start atomic structure, and 6DLV is the two-start low-resolution structure. * indicates assembly deficient mutant used in crystallographic studies. 3SNH: IHGIR395–399AAAAA; 3ZVR: G397D. b Schematic of dynamin 1 tetramer structures. The three conserved Stalk interfaces are denoted by numbers (1, 2, 3). c Typical SDS–PAGE (middle) and western blot (right) analysis of purified MBP-Dyn1ΔPRD. A primary antibody against dynamin 1 was used for the western blot. Three independent experiments were carried out. d Lipid-stimulated GTPase activity of MBP-Dyn1ΔPRD. Three independent experiments were carried out. std standard deviation. e–g Representative micrographs of negative-stained helical assemblies formed by MBP-Dyn1ΔPRD at 150 mM NaCl in the presence of GMP-PCP (e), DOPS (f), and both GMP-PCP and DOPS (g). h Immunogold labeling of MBP on MBP-Dyn1ΔPRD tubes shown in (e). Scale bars, 100 nm. e–h Three independent experiments were carried out.