Pheromone specificity of TraA and TraA-related proteins. The proteins were examined by SDS/PAGE and stained with silver stain. (A) Elution of amino terminus-fixed cAD1 affinity matrix-bound TraA-tag or TraA(1–310)-tag with various peptides. Protein extracts were mixed with the cAD1 matrix and, after washing, were eluted with 60 μM cAD1, iAD1, cPD1, iPD1, or cCF10. Each fraction (e1, e2, e3, and e4) represents 30 μl of eluate. (B) Amino terminus-fixed cAD1 and iAD1 affinity chromatography of TraA-tag, TraA(1–310)-tag, and TraA(1–300)-tag. Protein was eluted with 30 μl of 60 μM cAD1 or iAD1 in TSAGE buffer. In the case where protein was bound to the streptavidin matrix, elution was with 5 mM diaminobiotin.