Inactivation of Membrane Surface Ecto-5′-nucleotidase by Sodium Nitroprusside in C6 Glioma Cells
Abstract.: Ecto-5′-nucleotidase (NT5E), a predominant enzyme that produces extracellular adenosine from AMP, plays an important role in a variety of physiological and pathophysiological processes. This study was performed to identify agents that affect NT5E activity using C6 glioma cells. When cells...
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| Format: | Article |
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Elsevier
2011-01-01
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| Series: | Journal of Pharmacological Sciences |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S1347861319306590 |
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| author | Tadanori Kumasaka Isao Matsuoka Hirobumi Mashiko Shin-ichi Niwa Junko Kimura |
| author_facet | Tadanori Kumasaka Isao Matsuoka Hirobumi Mashiko Shin-ichi Niwa Junko Kimura |
| author_sort | Tadanori Kumasaka |
| collection | DOAJ |
| description | Abstract.: Ecto-5′-nucleotidase (NT5E), a predominant enzyme that produces extracellular adenosine from AMP, plays an important role in a variety of physiological and pathophysiological processes. This study was performed to identify agents that affect NT5E activity using C6 glioma cells. When cells were incubated with sodium nitroprusside (SNP), phorbol 12-myristate 13- acetate, forskolin, lipopolysaccharide, or interferon-γ, only SNP inhibited NT5E activity in a time- and concentration-dependent manner (IC50 = 1.2 μM). The inhibitory effect of SNP was long-lasting even after SNP washout; and its action was not mimicked by nitric oxide generating agents, 8-bromo cyclic GMP, ferricyanide, ferrocyanide, or sodium cyanide. SNP did not change NT5E mRNA level or membrane surface protein expression. Similar to SNP, Fe2+ inhibited NT5E activity, but to a lesser extent. Although Fe2+ is known to increase oxidative stress, Fe2+-mediated oxidative stress was not involved in SNP inhibition of NT5E because the inhibition of NT5E by SNP was not affected by superoxide dismutase and catalase. In contrast, addition of Zn2+, an essential metal co-factor of NT5E activity, prevented SNP from inhibiting NT5E. These results suggest that SNP disrupts a critical Zn2+-dependent enzyme activity and might be useful as a pharmacological tool for inhibiting NT5E. Keywords:: ecto-5′-nucleotidase, sodium nitroprusside, Zn2+, Fe2+, C6 glioma cell |
| first_indexed | 2024-12-10T07:56:24Z |
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| id | doaj.art-a74f1e9e6ce94d3f994b0e9dfb7f796a |
| institution | Directory Open Access Journal |
| issn | 1347-8613 |
| language | English |
| last_indexed | 2024-12-10T07:56:24Z |
| publishDate | 2011-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Pharmacological Sciences |
| spelling | doaj.art-a74f1e9e6ce94d3f994b0e9dfb7f796a2022-12-22T01:56:53ZengElsevierJournal of Pharmacological Sciences1347-86132011-01-0111714553Inactivation of Membrane Surface Ecto-5′-nucleotidase by Sodium Nitroprusside in C6 Glioma CellsTadanori Kumasaka0Isao Matsuoka1Hirobumi Mashiko2Shin-ichi Niwa3Junko Kimura4Department of Pharmacology, Fukushima 960-1295, Japan; Department of Neuropsychiatry, Fukushima Medical University School of Medicine, Fukushima 960-1295, JapanDepartment of Pharmacology, Fukushima 960-1295, Japan; Laboratory of Pharmacology, Faculty of Pharmacy, Takasaki University of Health and Welfare, Gunma 370-0033, Japan; Corresponding author (affiliation #3). isao@takasaki-u.ac.jpDepartment of Neuropsychiatry, Fukushima Medical University School of Medicine, Fukushima 960-1295, JapanDepartment of Neuropsychiatry, Fukushima Medical University School of Medicine, Fukushima 960-1295, JapanDepartment of Pharmacology, Fukushima 960-1295, JapanAbstract.: Ecto-5′-nucleotidase (NT5E), a predominant enzyme that produces extracellular adenosine from AMP, plays an important role in a variety of physiological and pathophysiological processes. This study was performed to identify agents that affect NT5E activity using C6 glioma cells. When cells were incubated with sodium nitroprusside (SNP), phorbol 12-myristate 13- acetate, forskolin, lipopolysaccharide, or interferon-γ, only SNP inhibited NT5E activity in a time- and concentration-dependent manner (IC50 = 1.2 μM). The inhibitory effect of SNP was long-lasting even after SNP washout; and its action was not mimicked by nitric oxide generating agents, 8-bromo cyclic GMP, ferricyanide, ferrocyanide, or sodium cyanide. SNP did not change NT5E mRNA level or membrane surface protein expression. Similar to SNP, Fe2+ inhibited NT5E activity, but to a lesser extent. Although Fe2+ is known to increase oxidative stress, Fe2+-mediated oxidative stress was not involved in SNP inhibition of NT5E because the inhibition of NT5E by SNP was not affected by superoxide dismutase and catalase. In contrast, addition of Zn2+, an essential metal co-factor of NT5E activity, prevented SNP from inhibiting NT5E. These results suggest that SNP disrupts a critical Zn2+-dependent enzyme activity and might be useful as a pharmacological tool for inhibiting NT5E. Keywords:: ecto-5′-nucleotidase, sodium nitroprusside, Zn2+, Fe2+, C6 glioma cellhttp://www.sciencedirect.com/science/article/pii/S1347861319306590 |
| spellingShingle | Tadanori Kumasaka Isao Matsuoka Hirobumi Mashiko Shin-ichi Niwa Junko Kimura Inactivation of Membrane Surface Ecto-5′-nucleotidase by Sodium Nitroprusside in C6 Glioma Cells Journal of Pharmacological Sciences |
| title | Inactivation of Membrane Surface Ecto-5′-nucleotidase by Sodium Nitroprusside in C6 Glioma Cells |
| title_full | Inactivation of Membrane Surface Ecto-5′-nucleotidase by Sodium Nitroprusside in C6 Glioma Cells |
| title_fullStr | Inactivation of Membrane Surface Ecto-5′-nucleotidase by Sodium Nitroprusside in C6 Glioma Cells |
| title_full_unstemmed | Inactivation of Membrane Surface Ecto-5′-nucleotidase by Sodium Nitroprusside in C6 Glioma Cells |
| title_short | Inactivation of Membrane Surface Ecto-5′-nucleotidase by Sodium Nitroprusside in C6 Glioma Cells |
| title_sort | inactivation of membrane surface ecto 5 nucleotidase by sodium nitroprusside in c6 glioma cells |
| url | http://www.sciencedirect.com/science/article/pii/S1347861319306590 |
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