Fluorescence spectrometry in studies of carbohydrate-protein interactions.
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Fluorometric spectroscopy is a powerful tool for investigating the interaction between a carbohydrate ligand and binding proteins. The measurement is done in situ and thus circumventing the need for separation of bound ligand from the free ligand. The source of the fluorophore can be intrinsic, i.e., the tryptophan in the protein, or extrinsic (contained in the ligand). Techniques for assessing the affinity used are measurement on fluorescence intensity change, lifetime, polarization anisotropy, and energy transfer. The last technique can also be used to study conformational structures of glycopeptides. It is also useful in designing substrates for endo-type enzymes which allow continuous monitoring of the reaction.