Journal: bioRxiv

Article Title: In vivo tests of the E. coli TonB system working model—interaction of ExbB with unknown proteins, identification of TonB-ExbD transmembrane heterodimers and PMF-dependent ExbD structures

doi: 10.1101/2024.07.10.602958

Figure Lengend Snippet: In strain RA1017, cultures expressing plasmid-encoded ExbB (pKP660), ExbB with a carboxy terminal T7 epitope tag (pKP735) and ExbB with an amino terminal His6 epitope tag (pKP560) were either DCCD-treated (+) or ethanol-treated (-), then formaldehyde-cross-linked and immunoblotted as in , except that a final concentration of 1 mM DCCD was used. Immunoblots were developed with anti-ExbB (upper panels) with the identities of the samples indicated across the top of the panel. The same samples were also developed with anti-T7 tag (bottom left panel) or anti-His tag antibodies (bottom right panel). The position of monomeric ExbB is indicated on the left. For ExbB from DCCD-treated samples, the yellow asterisk identifies ExbB monomer and red asterisk identifies ExbB L (lower). The two panels on the left show immunoblots from identical, divided samples developed with different antibodies; the two panels on the right show immunoblots from identical, divided samples developed with different antibodies.

Article Snippet: Anti-T7 tag-specific monoclonal antibodies were from Novagen.

Techniques: Expressing, Plasmid Preparation, Concentration Assay, Western Blot