STD-DOSY: A new NMR method to analyze multi-component enzyme/substrate systems.
A new approach to analyze multi-component Saturation Transfer Difference (STD) NMR spectra by combining the STD and the DOSY experiment is proposed. The resulting pulse sequence was successfully used to simplify an exemplary multi-component protein/substrate system by means of standard DOSY processing methods. Furthermore, the same experiment could be applied to calculate the ratio of saturated substrate molecules and its saturation rate in the case of competitive interactions. This ratio depends on the strength of this interaction between the substrates and the protein, so that this kind of information could be extracted from the results of our experiment.
Kramer
Markus
M
University of Potsdam, Department of Chemistry, Karl-Liebknecht-Str. 24-25, 14476 Potsdam/Golm, Germany.
Kleinpeter
Erich
E
D000465:Algorithms; D001665:Binding Sites; D003627:Data Interpretation, Statistical; D004058:Diffusion; D004798:Enzymes / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D011485:Protein Binding; D011506:Proteins / Q000737:chemistry; D013379:Substrate Specificity; D014909:Wheat Germ Agglutinins / Q000737:chemistry
D004798:Enzymes; D011506:Proteins; D014909:Wheat Germ Agglutinins