Figure 3.

Purification of recombinant EncFtn and Enc:EncFtn protein complexes. Recombinant EncFtn (A) and Enc:EncFtn (B) proteins were purified by anion exchange chromatography (Hi-Trap Q-Sepharose FF, GE Healthcare) and then subjected to size-exclusion chromatography using a Superdex 200 16/60 column (GE Healthcare) previously equilibrated with 50 mm Tris-HCl, pH 8.0, and 150 mm NaCl. The elution profiles in (A) reveal that all EncFtn proteins show a main peak at around 60 ml, diagnostic of oligomerization states close to 10-mer, and smaller peaks at ∼76 ml and 82 ml (indicative of smaller assembly states, Table S2). Enc:EncFtn protein complexes elute in a single peak around 46 ml, suggesting that EncFtn is compartmentalized within the encapsulin shell.