The stepwise binding of small molecules to proteins. Nuclear magnetic resonance and temperature jump studies of the binding of 4-(N-acetylaminoglucosyl)-N-acetylglucosamine to lysozyme.
The binding of 4-(N-acetylaminoglucosyl)-N-acetylglucosamine to lysozyme was studied by both nuclear magnetic resonance (NMR) and temperature-jump methods under comparable conditions. The NMR measurements on the inhibitor spectrum were carried out over a range of inhibitor concentrations including levels at which most of the inhibitor was bound to the enzyme. Data in this region were obtained by a novel difference method in conjunction with correlation spectroscopy. The results from the combination of both experimental techniques demonstrated the existence of a two-step binding mechanism and produced both values for all of the individual rate constants and also the NMR spectral data for the inhibitor in the two enzyme-inhibitor complexes. The later data characterize the environment experienced by the inhibitor at each stage in the binding process and thus provides both a three-dimensional and a dynamic picture of the interaction.
D000117:Acetylglucosamine; D000818:Animals; D001665:Binding Sites; D002645:Chickens; D004187:Disaccharides; D004529:Egg White; D005944:Glucosamine / Q000031:analogs & derivatives*; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D008433:Mathematics; D009113:Muramidase / Q000378:metabolism; D011485:Protein Binding; D011487:Protein Conformation; D013696:Temperature
D004187:Disaccharides; D009113:Muramidase; D005944:Glucosamine; D000117:Acetylglucosamine