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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MDPI AG
2021-01-01
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| Online Access: | https://www.mdpi.com/2073-4409/10/1/182 |
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| author | Rossana Pesi Simone Allegrini Francesco Balestri Mercedes Garcia-Gil Federico Cividini Laura Colombaioni Lars Petter Jordheim Marcella Camici Maria Grazia Tozzi |
| author_facet | Rossana Pesi Simone Allegrini Francesco Balestri Mercedes Garcia-Gil Federico Cividini Laura Colombaioni Lars Petter Jordheim Marcella Camici Maria Grazia Tozzi |
| author_sort | Rossana Pesi |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-03-09T04:25:11Z |
| format | Article |
| id | doaj.art-bd07450f4cf94e8781db2a3b7a186ba6 |
| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-09T04:25:11Z |
| publishDate | 2021-01-01 |
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| record_format | Article |
| series | Cells |
| spelling | doaj.art-bd07450f4cf94e8781db2a3b7a186ba62023-12-03T13:41:57ZengMDPI AGCells2073-44092021-01-0110118210.3390/cells10010182Cytosolic 5′-Nucleotidase II Is a Sensor of Energy Charge and Oxidative Stress: A Possible Function as Metabolic RegulatorRossana Pesi0Simone Allegrini1Francesco Balestri2Mercedes Garcia-Gil3Federico Cividini4Laura Colombaioni5Lars Petter Jordheim6Marcella Camici7Maria Grazia Tozzi8Unità di Biochimica, Dipartimento di Biologia, Università di Pisa, Via San Zeno 51, 56127 Pisa, ItalyUnità di Biochimica, Dipartimento di Biologia, Università di Pisa, Via San Zeno 51, 56127 Pisa, ItalyUnità di Biochimica, Dipartimento di Biologia, Università di Pisa, Via San Zeno 51, 56127 Pisa, ItalyUnità di Fisiologia Generale, Dipartimento di Biologia, Università di Pisa, Via San Zeno 31, 56127 Pisa, ItalyDepartment of Medicine, University of California, La Jolla, San Diego, CA 92093-0671, USAIstituto di Neuroscienze, CNR, Via Giuseppe Moruzzi 1, 56124 Pisa, ItalyCentre de Recherche en Cancérologie de Lyon, INSERM U1052-CNRS UMR 5286, Faculté Rockefeller, 8 Avenue Rockefeller, 69008 Lyon, FranceUnità di Biochimica, Dipartimento di Biologia, Università di Pisa, Via San Zeno 51, 56127 Pisa, ItalyUnità di Biochimica, Dipartimento di Biologia, Università di Pisa, Via San Zeno 51, 56127 Pisa, ItalyCytosolic 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.https://www.mdpi.com/2073-4409/10/1/182Cytosolic 5′-nucleotidase IINT5C2energy chargeoxidative stressAMPKADF |
| spellingShingle | Rossana Pesi Simone Allegrini Francesco Balestri Mercedes Garcia-Gil Federico Cividini Laura Colombaioni Lars Petter Jordheim Marcella Camici Maria Grazia Tozzi Cytosolic 5′-Nucleotidase II Is a Sensor of Energy Charge and Oxidative Stress: A Possible Function as Metabolic Regulator Cells Cytosolic 5′-nucleotidase II NT5C2 energy charge oxidative stress AMPK ADF |
| 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 | Cytosolic 5′-nucleotidase II NT5C2 energy charge oxidative stress AMPK ADF |
| url | https://www.mdpi.com/2073-4409/10/1/182 |
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