Fig. 1: Screening membrane scaffold peptides for detergent-free reconstitution of NDs.
From: DeFrND: detergent-free reconstitution into native nanodiscs with designer membrane scaffold peptides
a Illustration of the peptide screening strategy. Potent scaffold peptides will insert themselves into lipid bilayers of proteoliposomes and extract the maltose transporter MalFGK2 into NDs. Inset, surface charges of 18 A as an exemplary amphipathic scaffold peptide. The structural model of 18 A is generated by Alphafold2. MalF, blue; MalG, orange; MalK, yellow. b Representative blue-native gel analysis of detergent-free ND reconstitution. MalFGK2 proteoliposomes prepared using PC lipids were incubated with the indicated peptide scaffolds (33, 100 and 300 µM) in the absence of detergents and then analyzed by native electrophoresis. In control experiments, the same proteoliposomes were analyzed alone (lane 1) or treated with DDM (lane 2). Similar results were obtained with three independent experiments. c SEC profiles of detergent-free MalFGK2 NDs formed from proteoliposomes (pL) using 18 A, in comparison with control experiments of pL alone. Samples were fractionated on a Superdex S200 3.2/300 column. d ATPase activities of MalFGK2 in detergent micelles, pL, and 18A-enclosed NDs. Data are shown as mean ± s.d., n  =  3 independent experiments. e Negative stain EM micrograph of MalFGK2 NDs encased by 18 A. Red circles highlight the monomer NDs. Scale bar, 30 nm. Similar results were obtained with three independent experiments.
