Self-association of glucagon as measured by the optical properties of rhodamine 6G.
The fluorescence of rhodamine 6G is completely quenched in glucagon solutions in 0.6 M K2HOP4 at pH 10.6. The absorption of rhodamine 6G is red-shifted by the same reaction. A single rhodamine 6G molecule appears to be bound to a hydrophobic patch in the center of the trimer of glucagon. Since the glucagon monomer has almost no organized structure this site exists only in the associated trimer form of glucagon. The self-association of glucagon to the trimer has been determined from the variation in rhodamine 6G fluorescence and absorption measured over a 60-fold range of dye concentration. The self-association constant agrees with values determined by other methods in the absence of dye. The binding isotherms of rhodamine 6G to glucagon shift with glucagon concentration and exhibit negative cooperativity.
D000817:Anilino Naphthalenesulfonates; D005934:Glucagon; D007700:Kinetics; D046911:Macromolecular Substances; D011485:Protein Binding; D011487:Protein Conformation; D012235:Rhodamines; D013050:Spectrometry, Fluorescence; D014966:Xanthenes
D000817:Anilino Naphthalenesulfonates; D046911:Macromolecular Substances; D012235:Rhodamines; D014966:Xanthenes; D005934:Glucagon