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Pull-down of untagged EmrE by tagged EmrE-His. Membranes prepared from cells expressing tagged or untagged EmrE glycine mutants selectively labeled with [35S]methionine were solubilized in 0.8% DDM-Na buffer, mixed, and heat treated as described under “Experimental Procedures.” The tagged and untagged mixed proteins were then immobilized on Ni-NTA beads, washed, eluted, and separated by SDS-PAGE. Radioactive protein was visualized using a FLA-3000 PhosphorImager.
