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HMDC cross-linking of inactive glycine mutants. Membranes prepared from cells expressing EmrE glycine mutants selectively labeled with [35S]methionine were solubilized in 0.8% DDM-Na buffer, treated with HMDC as described under “Experimental Procedures,” and separated by SDS-PAGE. The lower bands (∼ 13 kDa) correspond to the monomer and the higher bands (∼26 kDa) to the cross-linked dimer. Radioactive protein was visualized using a FLA-3000 PhosphorImager.
