FIGURE 1.

Schematic illustration of the DirectMX methodology. The reconstitution of IMPs directly from crude cell membranes into Salipro nanoparticles is shown here for a recombinantly expressed example protein. (A) For DirectMX reconstitution of IMPs, homogenized crude cell membranes are incubated with digitonin and SapA. (B) The lipid-binding ability of SapA allows for self-assembly of native membrane lipid disks, thereby reconstituting IMPs into a scaffold of SapA molecules. (C) If the target IMP is fused to an affinity tag, the protein-containing nanoparticles can be readily purified by affinity chromatography using detergent-free buffers. The purified nanoparticles are then available for further downstream processes, such as SEC, or any kind of biochemical, structural or biophysical analysis.