Figure 6
From: [13C6,D8]2-deoxyglucose phosphorylation by hexokinase shows selectivity for the β-anomer
The β/α ratio of [13C6,D8]2DG and [13C6,D8]2DG6P over the course of the reaction with yHK, as calculated using signal deconvolution. (A) Spectra acquired at 3 time points from a typical experiment of [13C6,D8]2DG reaction in the presence of yHK (the same experiment is also shown in Figs. 2, 4, and 5). Blue lines indicate the position of signals indicative of [13C6,D8]2DG6P and red lines indicate position of signals indicative of [13C6,D8]2DG. For better visualization of the changes with time in the composition of the specific signals, the spectra at 14 s, and 22 s were multiplied 3-fold and 4-fold relative to the intensity of the spectrum at 2 s, respectively. (B) The ratio of product signal to total signal over time for the signals of the C1 and C6 positions in the same experiment as in (A). The rise in this ratio demonstrates the production of [13C6,D8]2DG6P. For the C6 position – this conclusion is based on the chemical shift difference of the substrate and the product signals as depicted in Fig. 2B. For the C1 position, this conclusion relies on the results of the deconvolution analysis of both anomers. The NRMSD between these two curves was 18%. (C) The β/α ratio of the C1 signal of [13C6,D8]2DG and [13C6,D8]2DG6P over time in the same experiment.
