Direct Quantitative Analysis of Enzyme-Catalyzed Reactions by Two-Dimensional Nuclear Magnetic Resonance Spectroscopy: Adenylate Kinase and Phosphoglyceromutase

Article abstract of DOI:10.1021/ja00261a027

The back-transformation method for the analysis of two-dimensional nuclear magnetic resonance cross-relaxation and exchange data has been employed to obtain the rate constants of the phosphoglyceromutase and adenylate kinase catalyzed reactions in vitro.The results are in excellent agreement with those obtained by other methods.It is shown that a single two-dimensional experiment for each catalyzed reaction is sufficient to calculate the rate constants.The study of the adenylate kinase reaction demonstrates the potential of the method for direct analysis of higher order reactions without having to simplify them in terms of pseudo-first-order steps.