Dyn2-Ser848 phosphorylation relieves Bin1 inhibition and promotes membrane fission. (A – C) Binding ability between Dyn2 and different SH3 domain. 12 µg GST, GST-Bin-SH3, or GST-endophilin-SH3 was incubated with 6 µg indicated purified Dyn2. Bound Dyn2 was analyzed with SDS-PAGE and Coomassie Blue staining. The ratio of bound Dyn2 was quantified, normalized to WT, and shown in B and C (n = 3 experimental replicates). Molecular weight is in kD. (D) Time-lapse representative images of Bin1-GFP tubules in cells co-expressing WT, S848A, or S848E Dyn2-mCherry were magnified and shown. White arrowheads indicate the occurrence of membrane fission. Scale bar, 2 µm. (E) Quantification of tubulated liposome diameter from coincubation of Bin1 and Dyn2 in Fig. 3 G (n ≥ 6 liposome tubules). (F) Multiple sequence alignment of Dyn2 PRD domain from different organisms. These sequences were analyzed with EMBL-EBI Clustal Omega Multiple Sequence Alignment. Data are shown as average ± SD and analyzed with one-way ANOVA. *P < 0.05; ***P < 0.001. Source data are available for this figure: SourceData FS2.