Fig. 1.

Membrane insertion of EHD2 requires ATP. (A) Structure representation of the domains and amino acids discussed in the manuscript based on the crystal structure of EHD2 (PDB ID code 4CID) (8, 9). The helical domain is depicted in purple, the flexible C-terminal EH domain in gray, and the core G domain in blue. (B) Adsorption of various EHD2 variants (wild type, EHD2 ΔEH, EHD2 ΔNΔEH, and EHD2 F122A/F128A) to lipid monolayers. The indicated proteins were incubated with an excess of AMP-PNP (solid lines) or ADP (dashed lines), respectively, before injection of the protein underneath the equilibrated lipid film (DOPC:POPS:PI(4,5)P2 = 70:27:3 mol) at time 0. The initial surface pressure of the lipid film (20–25 mN/m) was subtracted. The monolayer was stable over sufficient time for extensive IRRA spectroscopy.