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. 2019 Nov 14;8:e49787. doi: 10.7554/eLife.49787

Figure 5. The N-terminal strands of the substrate are assembled within the BamA β-barrel.

Figure 5.

(A) The lateral gate and interior of the BamA β-barrel interacts with the ends of the LptD substrate. The N-terminal strands of BamA (S439) interacts with the C-terminal strands of substrate LptD, while L6 (N666), within the β-barrel of BamA, interacts with the N-terminal strands of substrate LptD. MC4100 strains expressing a His-tagged BamA cysteine mutant and a FLAG-tagged LptD4213 cysteine mutant were treated with the cysteine-cysteine crosslinker 1,4-bismaleimidobutane (BMB). Crosslinked adducts of BamA and substrate LptD4213 were identified by immunoblot analyses after Ni-NTA affinity purification. Immunoblots are provided that show expression levels of cysteine-containing LptD4213 constructs from total cell lysates (bottom). (B) Map of cysteine crosslinking between the BamA and LptD4213. (C) Top-down cartoon representation of LptD engagement by BamA/D based on cysteine crosslinking data.