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| Content Provider | Springer Nature Link |
|---|---|
| Author | Qayyum, Imran Zubrow, Alan B. Ashraf, Qazi M. Kubin, Joanna Delivoria Papadopoulos, Maria Mishra, Om P. |
| Copyright Year | 2001 |
| Abstract | Previous studies have shown that hypoxia induces nitric oxide synthase-mediated generation of nitric oxide free radicals leading to peroxynitrite production. The present study tests the hypothesis that hypoxia results in NO-mediated modification of Na$^{+}$, K$^{+}$-ATPase in the fetal brain. Studies were conducted in guinea pig fetuses of 58-days gestation. The mothers were exposed to FiO$_{2}$ of 0.07% for 1 hour. Brain tissue hypoxia in the fetus was confirmed biochemically by decreased ATP and phosphocreatine levels. P$_{2}$ membrane fractions were prepared from normoxic and hypoxic fetuses and divided into untreated and treated groups. The membranes were treated with 0.5 mM peroxynitrite at pH 7.6. The Na$^{+}$, K$^{+}$-ATPase activity was determined at 37°C for five minutes in a medium containing 100 mM NaCl, 20 mM KCl, 6.0 mM MgCl$_{2}$, 50 mM Tris HCl buffer pH 7.4, 3.0 mM ATP with or without 10 mM ouabain. Ouabain sensitive activity was referred to as Na$^{+}$, K$^{+}$-ATPase activity. Following peroxynitrite exposure, the activity of Na$^{+}$, K$^{+}$-ATPase in guinea pig brain was reduced by 36% in normoxic membranes and further 29% in hypoxic membranes. Enzyme kinetics was determined at varying concentrations of ATP (0.5 mM-2.0 mM). The results indicate that peroxynitrite treatment alters the affinity of the active site of Na$^{+}$, K$^{+}$-ATPase for ATP and decreases the Vmax by 35% in hypoxic membranes. When compared to untreated normoxic membranes Vmax decreases by 35.6% in treated normoxic membranes and further to 52% in treated hypoxic membranes. The data show that peroxynitrite treatment induces modification of Na$^{+}$, K$^{+}$-ATPase. The results demonstrate that peroxynitrite decreased activity of Na$^{+}$, K$^{+}$-ATPase enzyme by altering the active sites as well as the microenvironment of the enzyme. We propose that nitric oxide synthase-mediated formation of peroxynitrite during hypoxia is a potential mechanism of hypoxia-induced decrease in Na$^{+}$, K$^{+}$-ATPase activity. |
| Starting Page | 1163 |
| Ending Page | 1169 |
| Page Count | 7 |
| File Format | |
| ISSN | 03643190 |
| Journal | Neurochemical Research |
| Volume Number | 26 |
| Issue Number | 10 |
| e-ISSN | 15736903 |
| Language | English |
| Publisher | Kluwer Academic Publishers-Plenum Publishers |
| Publisher Date | 2001-01-01 |
| Publisher Place | New York |
| Access Restriction | One Nation One Subscription (ONOS) |
| Subject Keyword | Neurosciences Neurology Biochemistry |
| Content Type | Text |
| Resource Type | Article |
| Subject | Medicine Biochemistry Cellular and Molecular Neuroscience |
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