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Real-Time Pyruvate Chemical Conversion Monitoring Enabled by PHIP

MPG-Autoren
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Stevanato,  Gabriele
Research Group of NMR Signal Enhancement, Max Planck Institute for Multidisciplinary Sciences, Max Planck Society;

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Ding,  Yonghong
Research Group of NMR Signal Enhancement, Max Planck Institute for Multidisciplinary Sciences, Max Planck Society;

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Mamone,  Salvatore
Research Group of NMR Signal Enhancement, Max Planck Institute for Multidisciplinary Sciences, Max Planck Society;

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Jagtap,  Anil P.
Research Group of NMR Signal Enhancement, Max Planck Institute for Multidisciplinary Sciences, Max Planck Society;

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Korchak,  Sergey
Research Group of NMR Signal Enhancement, Max Planck Institute for Multidisciplinary Sciences, Max Planck Society;

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Glöggler,  Stefan
Research Group of NMR Signal Enhancement, Max Planck Institute for Multidisciplinary Sciences, Max Planck Society;

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Zitation

Stevanato, G., Ding, Y., Mamone, S., Jagtap, A. P., Korchak, S., & Glöggler, S. (2023). Real-Time Pyruvate Chemical Conversion Monitoring Enabled by PHIP. Journal of the American Chemical Society, 145(10), 5864-5871. doi:10.1021/jacs.2c13198.


Zitierlink: https://hdl.handle.net/21.11116/0000-000C-CAD4-1
Zusammenfassung
In recent years, parahydrogen-induced polarization side arm hydrogenation (PHIP-SAH) has been applied to hyperpolarize [1-13C]pyruvate and map its metabolic conversion to [1-13C]lactate in cancer cells. Developing on our recent MINERVA pulse sequence protocol, in which we have achieved 27% [1-13C]pyruvate carbon polarization, we demonstrate the hyperpolarization of [1,2-13C]pyruvate (∼7% polarization on each 13C spin) via PHIP-SAH. By altering a single parameter in the pulse sequence, MINERVA enables the signal enhancement of C1 and/or C2 in [1,2-13C]pyruvate with the opposite phase, which allows for the simultaneous monitoring of different chemical reactions with enhanced spectral contrast or for the same reaction via different carbon sites. We first demonstrate the ability to monitor the same enzymatic pyruvate to lactate conversion at 7T in an aqueous solution, in vitro, and in-cell (HeLa cells) via different carbon sites. In a second set of experiments, we use the C1 and C2 carbon positions as spectral probes for simultaneous chemical reactions: the production of acetate, carbon dioxide, bicarbonate, and carbonate by reacting [1,2-13C]pyruvate with H2O2 at a high temperature (55 °C). Importantly, we detect and characterize the intermediate 2-hydroperoxy-2-hydroxypropanoate in real time and at high temperature.