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. 2009 Feb 20;284(8):5403–5413. doi: 10.1074/jbc.M806012200

FIGURE 2.

FIGURE 2.

Intact RseB is required for complete binding to RseA. SDS-PAGE analysis of in vitro binding of His-tagged RseA-peri to Flag-tagged RseB (A) or Flag-tagged RseB C-terminal domain (B); and of His-Trx-tagged RseA 169-196 peptide to Flag-tagged RseB (D) or Flag-tagged RseB C-terminal domain (E). RseA was immobilized on Ni-NTA resin and was allowed to interact with an equimolar amount of RseB (shown as input, In). Unbound protein was collected as flow through (F/T), bound proteins were eluted with imidazole (eluate, E) after washing. C and F presents relative binding of RseB to RseA [(wt RseB or RseB C-terminal domain/RseA)/(wt RseB/RseA) × 100] after quantification of data with ImageJ software as described in Fig. 1 and “Experimental Procedures.” These experiments were repeated three times, and similar results were obtained.