Fig. 5. The conformational changes of the Plug-domain in different P5A states and mass-spectrometry-identified possible cargo.

a Non-sharpened cryo-EM map of the here determined states. The Plug-domain is highlighted in green and the CtSpf1 and the nanodiscs are in gray. b Close-views displaying the conformational changes of the modeled parts of the Plug-domain (green), the connection to the ATPase core (blue), and the TM domain (orange and wheat). c Overview table of proteins with at least one TM helix (including signal peptides, SP) identified with MS. Hits exclusively found in CtSpf1 in the presence of BeF₃^- and not in controls, and hits among the 20 most overrepresented in the same CtSpf1 (BeF₃⁻) sample compared to GFP in the presence of BeF₃⁻ have been included. Two SPs were identified, both exclusively found in CtSpf1(BeF₃⁻). The table distinguishes between identified proteins with one or multiple TM helices (for a detailed list of these hits, see Supplementary Table 3), and specifies the in-protein location of the TM helix in the identified 1 TM proteins. It is likely that some of these hits represent transport cargo for CtSpf1. d Overview of the identified proteins containing an SP or one TM helix, displaying the overall topology (when correctly inserted), the number of identified hits, and the membrane into which the hits are inserted. TA denotes non-mitochondrial TA proteins. For a complete list of the proteins, see Supplementary Table 3. Figure 5d was generated using BioRender; https://BioRender.com/n26e116 - Gourdon, P. (2024).