Fig. 5.

ATP hydrolysis abolishes spontaneous transfer of PLs from MlaFEDB to MlaC. (A) SDS-PAGE analysis of indicated samples following coincubation with or without ATP, or vanadate, and subsequent nickel affinity–based clean separation of tagless nonradioactive apo MlaC (10 μM) and His-tagged nanodisc-embedded complexes reconstituted with [¹⁴C]-PLs (2 μM). !, apo MlaC; #, nanodisc-embedded [¹⁴C]-MlaFEDB; %, nanodisc-embedded [¹⁴C]-MlaFEB; SS, silver staining. (B) Average gains and losses of radioactivity ([¹⁴C]-lipids) in MlaC (Top) and MlaFEB (Bottom), respectively, normalized to the initial counts on MlaFEB. (C) Average gains and losses of radioactivity ([¹⁴C]-lipids) in MlaC (Top) and MlaFEDB (Bottom), respectively, normalized to the initial counts on MlaFEDB. Data from coincubation of MlaFEB/MlaFEDB with nickel resin alone are treated as unspecific background loss. Error bars represent SDs calculated from triplicate experiments. Student’s t tests: ns, not significant; *P < 0.05; ****P < 0.0001.