Figure 5.

TEV protease cleavage of the N-terminal expression tag MGSSHHHHHHSSGENLYFQ in [80%-²H,u-¹⁵N]-His₆-TEV-OmpA reconstituted in DMPC nanodiscs. A) SDS PAGE. 1: Reaction mixture before addition of the TEV protease; 2 and 3: Reaction mixture after incubation at 4°C for 24 h and 48 h, respectively, following the addition of the TEV protease; MWM: Molecular weight markers. Assignments of the protein bands are indicated on the left. Arrows highlight the position of cleaved OmpA, which is partially overlapped with the scaffold protein band. B) Overlay of the 2D [¹⁵N,¹H]-TROSY correlation NMR spectra of OmpA in DMPC nanodiscs purified before (red) and after (blue) TEV protease cleavage for 48 h at 4 °C.