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Cytosolic 5′-Nucleotidase II Is a Sensor of Energy Charge and Oxidative Stress: A Possible Function as Metabolic Regulator

Cytosolic 5′-nucleotidase II (NT5C2) is a highly regulated enzyme involved in the maintenance of intracellular purine and the pyrimidine compound pool. It dephosphorylates mainly IMP and GMP but is also active on AMP. This enzyme is highly expressed in tumors, and its activity correlates with a high...

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Autores principales: Pesi, Rossana, Allegrini, Simone, Balestri, Francesco, Garcia-Gil, Mercedes, Cividini, Federico, Colombaioni, Laura, Jordheim, Lars Petter, Camici, Marcella, Tozzi, Maria Grazia
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7831490/
https://www.ncbi.nlm.nih.gov/pubmed/33477638
http://dx.doi.org/10.3390/cells10010182
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author Pesi, Rossana
Allegrini, Simone
Balestri, Francesco
Garcia-Gil, Mercedes
Cividini, Federico
Colombaioni, Laura
Jordheim, Lars Petter
Camici, Marcella
Tozzi, Maria Grazia
author_facet Pesi, Rossana
Allegrini, Simone
Balestri, Francesco
Garcia-Gil, Mercedes
Cividini, Federico
Colombaioni, Laura
Jordheim, Lars Petter
Camici, Marcella
Tozzi, Maria Grazia
author_sort Pesi, Rossana
collection PubMed
description Cytosolic 5′-nucleotidase II (NT5C2) is a highly regulated enzyme involved in the maintenance of intracellular purine and the pyrimidine compound pool. It dephosphorylates mainly IMP and GMP but is also active on AMP. This enzyme is highly expressed in tumors, and its activity correlates with a high rate of proliferation. In this paper, we show that the recombinant purified NT5C2, in the presence of a physiological concentration of the inhibitor inorganic phosphate, is very sensitive to changes in the adenylate energy charge, especially from 0.4 to 0.9. The enzyme appears to be very sensitive to pro-oxidant conditions; in this regard, the possible involvement of a disulphide bridge (C175-C547) was investigated by using a C547A mutant NT5C2. Two cultured cell models were used to further assess the sensitivity of the enzyme to oxidative stress conditions. NT5C2, differently from other enzyme activities, was inactivated and not rescued by dithiothreitol in a astrocytoma cell line (ADF) incubated with hydrogen peroxide. The incubation of a human lung carcinoma cell line (A549) with 2-deoxyglucose lowered the cell energy charge and impaired the interaction of NT5C2 with the ice protease-activating factor (IPAF), a protein involved in innate immunity and inflammation.
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spelling pubmed-78314902021-01-26 Cytosolic 5′-Nucleotidase II Is a Sensor of Energy Charge and Oxidative Stress: A Possible Function as Metabolic Regulator Pesi, Rossana Allegrini, Simone Balestri, Francesco Garcia-Gil, Mercedes Cividini, Federico Colombaioni, Laura Jordheim, Lars Petter Camici, Marcella Tozzi, Maria Grazia Cells Article Cytosolic 5′-nucleotidase II (NT5C2) is a highly regulated enzyme involved in the maintenance of intracellular purine and the pyrimidine compound pool. It dephosphorylates mainly IMP and GMP but is also active on AMP. This enzyme is highly expressed in tumors, and its activity correlates with a high rate of proliferation. In this paper, we show that the recombinant purified NT5C2, in the presence of a physiological concentration of the inhibitor inorganic phosphate, is very sensitive to changes in the adenylate energy charge, especially from 0.4 to 0.9. The enzyme appears to be very sensitive to pro-oxidant conditions; in this regard, the possible involvement of a disulphide bridge (C175-C547) was investigated by using a C547A mutant NT5C2. Two cultured cell models were used to further assess the sensitivity of the enzyme to oxidative stress conditions. NT5C2, differently from other enzyme activities, was inactivated and not rescued by dithiothreitol in a astrocytoma cell line (ADF) incubated with hydrogen peroxide. The incubation of a human lung carcinoma cell line (A549) with 2-deoxyglucose lowered the cell energy charge and impaired the interaction of NT5C2 with the ice protease-activating factor (IPAF), a protein involved in innate immunity and inflammation. MDPI 2021-01-18 /pmc/articles/PMC7831490/ /pubmed/33477638 http://dx.doi.org/10.3390/cells10010182 Text en © 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Pesi, Rossana
Allegrini, Simone
Balestri, Francesco
Garcia-Gil, Mercedes
Cividini, Federico
Colombaioni, Laura
Jordheim, Lars Petter
Camici, Marcella
Tozzi, Maria Grazia
Cytosolic 5′-Nucleotidase II Is a Sensor of Energy Charge and Oxidative Stress: A Possible Function as Metabolic Regulator
title Cytosolic 5′-Nucleotidase II Is a Sensor of Energy Charge and Oxidative Stress: A Possible Function as Metabolic Regulator
title_full Cytosolic 5′-Nucleotidase II Is a Sensor of Energy Charge and Oxidative Stress: A Possible Function as Metabolic Regulator
title_fullStr Cytosolic 5′-Nucleotidase II Is a Sensor of Energy Charge and Oxidative Stress: A Possible Function as Metabolic Regulator
title_full_unstemmed Cytosolic 5′-Nucleotidase II Is a Sensor of Energy Charge and Oxidative Stress: A Possible Function as Metabolic Regulator
title_short Cytosolic 5′-Nucleotidase II Is a Sensor of Energy Charge and Oxidative Stress: A Possible Function as Metabolic Regulator
title_sort cytosolic 5′-nucleotidase ii is a sensor of energy charge and oxidative stress: a possible function as metabolic regulator
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7831490/
https://www.ncbi.nlm.nih.gov/pubmed/33477638
http://dx.doi.org/10.3390/cells10010182
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