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Excess exogenous pyruvate inhibits lactate dehydrogenase activity in live cells in an MCT1-dependent manner

Cellular pyruvate is an essential metabolite at the crossroads of glycolysis and oxidative phosphorylation, capable of supporting fermentative glycolysis by reduction to lactate mediated by lactate dehydrogenase (LDH) among other functions. Several inherited diseases of mitochondrial metabolism impa...

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Autores principales: Rao, Yi, Gammon, Seth T., Sutton, Margie N., Zacharias, Niki M., Bhattacharya, Pratip, Piwnica-Worms, David
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society for Biochemistry and Molecular Biology 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8233206/
https://www.ncbi.nlm.nih.gov/pubmed/34022218
http://dx.doi.org/10.1016/j.jbc.2021.100775
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author Rao, Yi
Gammon, Seth T.
Sutton, Margie N.
Zacharias, Niki M.
Bhattacharya, Pratip
Piwnica-Worms, David
author_facet Rao, Yi
Gammon, Seth T.
Sutton, Margie N.
Zacharias, Niki M.
Bhattacharya, Pratip
Piwnica-Worms, David
author_sort Rao, Yi
collection PubMed
description Cellular pyruvate is an essential metabolite at the crossroads of glycolysis and oxidative phosphorylation, capable of supporting fermentative glycolysis by reduction to lactate mediated by lactate dehydrogenase (LDH) among other functions. Several inherited diseases of mitochondrial metabolism impact extracellular (plasma) pyruvate concentrations, and [1-(13)C]pyruvate infusion is used in isotope-labeled metabolic tracing studies, including hyperpolarized magnetic resonance spectroscopic imaging. However, how these extracellular pyruvate sources impact intracellular metabolism is not clear. Herein, we examined the effects of excess exogenous pyruvate on intracellular LDH activity, extracellular acidification rates (ECARs) as a measure of lactate production, and hyperpolarized [1-(13)C]pyruvate-to-[1-(13)C]lactate conversion rates across a panel of tumor and normal cells. Combined LDH activity and LDHB/LDHA expression analysis intimated various heterotetrameric isoforms comprising LDHA and LDHB in tumor cells, not only canonical LDHA. Millimolar concentrations of exogenous pyruvate induced substrate inhibition of LDH activity in both enzymatic assays ex vivo and in live cells, abrogated glycolytic ECAR, and inhibited hyperpolarized [1-(13)C]pyruvate-to-[1-(13)C]lactate conversion rates in cellulo. Of importance, the extent of exogenous pyruvate-induced inhibition of LDH and glycolytic ECAR in live cells was highly dependent on pyruvate influx, functionally mediated by monocarboxylate transporter-1 localized to the plasma membrane. These data provided evidence that highly concentrated bolus injections of pyruvate in vivo may transiently inhibit LDH activity in a tissue type- and monocarboxylate transporter-1–dependent manner. Maintaining plasma pyruvate at submillimolar concentrations could potentially minimize transient metabolic perturbations, improve pyruvate therapy, and enhance quantification of metabolic studies, including hyperpolarized [1-(13)C]pyruvate magnetic resonance spectroscopic imaging and stable isotope tracer experiments.
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spelling pubmed-82332062021-06-29 Excess exogenous pyruvate inhibits lactate dehydrogenase activity in live cells in an MCT1-dependent manner Rao, Yi Gammon, Seth T. Sutton, Margie N. Zacharias, Niki M. Bhattacharya, Pratip Piwnica-Worms, David J Biol Chem Research Article Cellular pyruvate is an essential metabolite at the crossroads of glycolysis and oxidative phosphorylation, capable of supporting fermentative glycolysis by reduction to lactate mediated by lactate dehydrogenase (LDH) among other functions. Several inherited diseases of mitochondrial metabolism impact extracellular (plasma) pyruvate concentrations, and [1-(13)C]pyruvate infusion is used in isotope-labeled metabolic tracing studies, including hyperpolarized magnetic resonance spectroscopic imaging. However, how these extracellular pyruvate sources impact intracellular metabolism is not clear. Herein, we examined the effects of excess exogenous pyruvate on intracellular LDH activity, extracellular acidification rates (ECARs) as a measure of lactate production, and hyperpolarized [1-(13)C]pyruvate-to-[1-(13)C]lactate conversion rates across a panel of tumor and normal cells. Combined LDH activity and LDHB/LDHA expression analysis intimated various heterotetrameric isoforms comprising LDHA and LDHB in tumor cells, not only canonical LDHA. Millimolar concentrations of exogenous pyruvate induced substrate inhibition of LDH activity in both enzymatic assays ex vivo and in live cells, abrogated glycolytic ECAR, and inhibited hyperpolarized [1-(13)C]pyruvate-to-[1-(13)C]lactate conversion rates in cellulo. Of importance, the extent of exogenous pyruvate-induced inhibition of LDH and glycolytic ECAR in live cells was highly dependent on pyruvate influx, functionally mediated by monocarboxylate transporter-1 localized to the plasma membrane. These data provided evidence that highly concentrated bolus injections of pyruvate in vivo may transiently inhibit LDH activity in a tissue type- and monocarboxylate transporter-1–dependent manner. Maintaining plasma pyruvate at submillimolar concentrations could potentially minimize transient metabolic perturbations, improve pyruvate therapy, and enhance quantification of metabolic studies, including hyperpolarized [1-(13)C]pyruvate magnetic resonance spectroscopic imaging and stable isotope tracer experiments. American Society for Biochemistry and Molecular Biology 2021-05-20 /pmc/articles/PMC8233206/ /pubmed/34022218 http://dx.doi.org/10.1016/j.jbc.2021.100775 Text en © 2021 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Research Article
Rao, Yi
Gammon, Seth T.
Sutton, Margie N.
Zacharias, Niki M.
Bhattacharya, Pratip
Piwnica-Worms, David
Excess exogenous pyruvate inhibits lactate dehydrogenase activity in live cells in an MCT1-dependent manner
title Excess exogenous pyruvate inhibits lactate dehydrogenase activity in live cells in an MCT1-dependent manner
title_full Excess exogenous pyruvate inhibits lactate dehydrogenase activity in live cells in an MCT1-dependent manner
title_fullStr Excess exogenous pyruvate inhibits lactate dehydrogenase activity in live cells in an MCT1-dependent manner
title_full_unstemmed Excess exogenous pyruvate inhibits lactate dehydrogenase activity in live cells in an MCT1-dependent manner
title_short Excess exogenous pyruvate inhibits lactate dehydrogenase activity in live cells in an MCT1-dependent manner
title_sort excess exogenous pyruvate inhibits lactate dehydrogenase activity in live cells in an mct1-dependent manner
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8233206/
https://www.ncbi.nlm.nih.gov/pubmed/34022218
http://dx.doi.org/10.1016/j.jbc.2021.100775
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