Figure 5
From: [13C6,D8]2-deoxyglucose phosphorylation by hexokinase shows selectivity for the β-anomer
Signal deconvolution of the C1β and C1α signals to their respective [13C6,D8]2DG and [13C6,D8]2DG6P components in one spectrum from the reaction of [13C6,D8]2DG with yHK. This specific spectrum was acquired 14 s after the beginning of the reaction with yHK. The spectrum is taken from the same experiment that is shown in Fig. 2B,A. (A) The fitted signal components of C1β and C1α of [13C6,D8]2DG. The complex signal of each anomer consists of 6 Lorentzian lines due to D-13C coupling (J13C-D = 25.5 Hz) and 13C-13C coupling (J13C-13C = 39.7 Hz). The J-coupling constants reported here are in agreement with those previously reported for [13C6,D7]glucose50 and for [1–13C]glucose77. The 13C-13C doublet component of each signal was allowed to be asymmetric as asymmetry was previously observed in 13C-13C split signals in a hyperpolarized state (and occasionally at thermal equilibrium)38,78,79. Both fit components are shown in solid light green line, the sum of both components is shown in dashed dark green line. (B) The fitted signal components of C1β and C1α of [13C6,D8]2DG6P. The complex signal of each anomer consists of 6 Lorentzian lines due to D-13C coupling (J13C-D = 25.1 Hz) and due to 13C-13C coupling (J13C-13C = 38.6 Hz). The 13C-13C doublet component of each signal was allowed to be asymmetric here as well. Both fit components are shown in solid cyan line, the sum of both components is shown in the solid blue line. (C) The combination of the 2DG and the 2DG6P fitted signals components. The sum of 2DG fit components is shown in the dashed dark green line, the sum of 2DG6P fit components is shown in the solid blue line, and the sum of both 2DG and 2DG6P fit components is shown in the black dotted line. (D) A comparison between the overall fitted signals (black dotted line) shown in (C) to the experimental spectrum (solid black line) demonstrates the utility of the deconvolution procedure. The normalized root mean square deviation (NRMSD) between the fit and the experimentally measured spectrum was found to be 9% and 17% for the C1β and C1α anomers, respectively. 2DG, [13C6,D8]2DG; 2DG6P, [13C6,D8]2DG6P.
