Adsorption isotherms of ouabain on hepatocytes from normal and diabetic (streptozotocin-induced) rats.
A cell surface adsorption isotherm approach is investigated with normal and diabetic (streptozotocin-induced) rat hepatocytes utilizing mathematical modeling. Freshly prepared monodispersed viable rat hepatocytes in Ca(2+)- and Mg(2+)-free phosphate buffer are obtained by collagenase perfusion and used in this study. [3H]ouabain is used as a ligand that specifically binds with the alpha 1 and alpha 2 isoforms of the alpha-protein subunit of the hepatocyte-membrane-incorporated Na-K-ATPase. The model that fits the experimental data assumes the presence of multiple receptors on the cell surface, and only when a specific fraction of the total number of one receptor have effectively reacted will the other receptor initiate reaction with the ligand. The results suggest the existence of two receptors, in normal and diabetic hepatocytes, interacting with ouabain and having different equilibrium constants. The alpha 2 isoform interacts more strongly with ouabain than the alpha 1 isoform in both types of cells. The alpha 1 isoform of the diabetic hepatocytes has stronger affinity with the glycoside than the alpha 1 isoform of the normal hepatocytes, while alpha 2 of the diabetics shows weaker affinity than alpha 2 of the normal hepatocytes. Therefore, the alpha 1 and alpha 2 isoforms of Na-K-ATPase in hepatocyte-cell-membrane have different affinities for ouabain and have been conformationally and/or structurally altered in chronic diabetes.
Bikhazi
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Department of Physiology, American University of Beirut, New York, NY 10022.
Bitar
K M
KM
Shaaban
E K
EK
el-Kasti
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MM
D000327:Adsorption; D000818:Animals; D003921:Diabetes Mellitus, Experimental / Q000378:metabolism*; D007700:Kinetics; D008099:Liver / Q000166:cytology / Q000378:metabolism*; D046911:Macromolecular Substances; D008297:Male; D008433:Mathematics; D010042:Ouabain / Q000378:metabolism / Q000493:pharmacokinetics*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012680:Sensitivity and Specificity; D000254:Sodium-Potassium-Exchanging ATPase / Q000378:metabolism; D014316:Tritium
D046911:Macromolecular Substances; D014316:Tritium; D010042:Ouabain; D000254:Sodium-Potassium-Exchanging ATPase