Absence of 633-nm laser irradiation-induced effects on glucose phosphorylation by hexokinase

In a paper by Amat et al. (Modification of the intrinsic fluorescence and biochemical behavior of adenosine triphosphate ATP after irradiation with visible and near-infrared laser light, J. Photochem. Photobiol. B 81 (2005) 26-32) it was shown that the conversion of glucose to glucose-6-phosphate by hexokinase in vitro was accelerated when ATP, which supplies the reaction with energy, was priorly irradiated at non-resonant optical frequencies (NROF, i.e., 655 and 830 nm). Correspondingly, the authors postulated that NROF may lower the energy barrier for the dephosphorylation of ATP's terminal phosphate and thus accelerate the reaction rate through a more expeditious energy delivery. Next to the established photobiostimulatory influence of visible light on cells, which is mediated by cytochrome c oxidase through resonant effects of light, Amat et al. posited an interesting theory with which the same processes could be induced through non-resonant effects. To investigate the effects of NROF with respect to the hexokinase reaction in greater detail, the reaction rates were measured spectrofluorometrically after 633-nm laser irradiation of ATP, the ATP-Mg complex, hexokinase, and the entire reaction mixture at room temperature (22 degrees C) and at the optimal reaction temperature (30 degrees C). No differences in reaction rates between the NROF-irradiated and control groups were found at either temperature. The hypothesis that NROF enhances in vitro hexokinase activity by lowering the activation energy for the dephosphorylation of ATP's terminal phosphate by hexokinase was therefore disproven. Consequently, it is questionable, albeit not unequivocal, that NROF exerts an effect on other ATP-driven reactions in cell metabolic pathways through a direct impact on ATP.