Extended Data Figure 2. Role of P294 in BSE conformational dynamics.
A. Changes in emission intensity of BODIPY-labeled CW-Dyn1 and P294A-CW-Dyn1 due to loss of PET following addition of 1 mM GMPPCP. Although the BSE partially opens upon addition of GMPPCP, its movements are constrained relative to WT by the mutation of P294. B. Assembly-stimulated GTPase activity of 0.5 μM P294A, P294G and P294V-Dyn1 measured on 100 nm liposomes relative to WT-Dyn1. The mutants show near WT activity indicating their ability to self-assemble onto and tubulate liposomes (data shown as average ±SD, n=4). C. Fission activity of 0.5 μM P294A, P294G and P294V-Dyn1 relative to WT-Dyn1 measured as the percentage of total membrane released from SUPER templates during 30 min incubation in the presence of GTP (Data shown as average ±SD, n=3). Substitution of P294 with the more rigid valine residue has a greater effect on fission activity.