Pyruvate cellular uptake and enzymatic conversion probed by dissolution DNP‐NMR: the impact of overexpressed membrane transporters |
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| Authors: | Riccardo Balzan Laetitia Fernandes Laetitia Pidial Arnaud Comment Bertrand Tavitian Paul R Vasos |
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| Institution: | 1. Laboratoire de Chimie et Biochimie Pharmacologiques et Toxicologiques, UMR 8601, UFR Biomédicale et des Sciences de Base, Université Paris Descartes – CNRS, Paris, France;2. PARCC – Inserm U970 – Faculté de Médecine, Université Paris Descartes, Paris, France;3. EPFL, Institute of Physics of Biological Systems, Lausanne, Switzerland |
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| Abstract: | Pyruvate membrane crossing and its lactate dehydrogenase‐mediated conversion to lactate in cells featuring different levels of expression of membrane monocarboxylate transporters (MCT4) were probed by dissolution dynamic nuclear polarization‐enhanced NMR. Hyperpolarized 13C‐1‐labeled pyruvate was transferred to suspensions of rodent tumor cell carcinoma, cell line 39. The pyruvate‐to‐lactate conversion rate monitored by dissolution dynamic nuclear polarization‐NMR in carcinoma cells featuring native MCT4 expression level was lower than the rate observed for cells in which the human MCT4 gene was overexpressed. The enzymatic activity of lactate dehydrogenase was also assessed in buffer solutions, following the real‐time pyruvate‐to‐lactate conversion speeds at different enzyme concentrations. Copyright © 2016 John Wiley & Sons, Ltd. |
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| Keywords: | dissolution dynamic nuclear polarization (d‐DNP) nuclear magnetic resonance (NMR) lactate dehydrogenase (LDH) rodent cancer cells metabolism |
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