Applications of blue dextran and cibacron blue F3GA in purification and structural studies of nucleotide-requiring enzymes

Abstract

Summary Cibacron Blue F3GA, the chromophore of Blue Dextran, interacted with all eight nucleotide-requiring enzymes that were examined. The conjugated chromophore (Blue Dextran) was more selective, interacting with enzymes including (but perhaps not restricted to) those known to possess the “dinucleotide fold”. The discriminating ability of these ligands should prove useful in elucidating the nature of nucleotide binding sites, and facilitate purification of selected nucleotide-requiring enzymes using affinity chromatography methods. Based on their comparable interactions with Cibacron Blue F3GA and weak interactions with Blue Dextran, it is suggested that neither yeast nor rat brain hexokinase contains the dinucleotide fold as a structural feature.