[3H]ouabain binding to cultured endothelial cells: effect of cholesterol enrichment. 1994

Y T Lau, and J K Chen, and M M Hsu, and H M Yu
Department of Physiology, Chang Gung Medical College, Kweisan, Taiwan, ROC.

Binding experiments were performed with [3H]ouabain on cultured human umbilical vein endothelial cells (huvEC). Saturation studies yielded a binding capacity (Bmax) of 820 +/- 81 fmole/mg pr.(n = 4) and dissociation constant (KD) of 11.7 +/- 2.1nM (n = 4) in K(+)-free buffer for specific [3H] ouabain binding on these cells. External K+ inhibited this binding in a dose-dependent manner. The mean value of Bmax is equivalent to about 4 x 10(5) sites per cell, comparable with that of smooth muscle cell. These data demonstrated the presence of specific [3H]ouabain binding linked to Na+/K+ pump, consistent with the observations of ouabain-sensitive 86Rb uptake in huvEC. Effect of cholesterol enrichment was also studied. Incubation in media supplemented with cholesterol-phospholipid liposomes of molar ratio of 2:1 for 18 hours reduced the Bmax by 31% (P < 0.05) without significantly changed the value of KD. This reduction of [3H]ouabain binding appeared to be specific for cholesterol since liposome made with pure phospholipid did not alter binding. Recent findings indicate that cholesterol-enrichment and plasma lipoproteins enhance vascular contractile response, our results suggest that the cholesterol-enrichment of endothelial cells may also indirectly affect the vascular response via disturbing the function of Na+/K+ pump.

UI MeSH Term Description Entries
D008081 Liposomes Artificial, single or multilaminar vesicles (made from lecithins or other lipids) that are used for the delivery of a variety of biological molecules or molecular complexes to cells, for example, drug delivery and gene transfer. They are also used to study membranes and membrane proteins. Niosomes,Transferosomes,Ultradeformable Liposomes,Liposomes, Ultra-deformable,Liposome,Liposome, Ultra-deformable,Liposome, Ultradeformable,Liposomes, Ultra deformable,Liposomes, Ultradeformable,Niosome,Transferosome,Ultra-deformable Liposome,Ultra-deformable Liposomes,Ultradeformable Liposome
D010042 Ouabain A cardioactive glycoside consisting of rhamnose and ouabagenin, obtained from the seeds of Strophanthus gratus and other plants of the Apocynaceae; used like DIGITALIS. It is commonly used in cell biological studies as an inhibitor of the NA(+)-K(+)-EXCHANGING ATPASE. Acocantherin,G-Strophanthin,Acolongifloroside K,G Strophanthin
D011188 Potassium An element in the alkali group of metals with an atomic symbol K, atomic number 19, and atomic weight 39.10. It is the chief cation in the intracellular fluid of muscle and other cells. Potassium ion is a strong electrolyte that plays a significant role in the regulation of fluid volume and maintenance of the WATER-ELECTROLYTE BALANCE.
D002478 Cells, Cultured Cells propagated in vitro in special media conducive to their growth. Cultured cells are used to study developmental, morphologic, metabolic, physiologic, and genetic processes, among others. Cultured Cells,Cell, Cultured,Cultured Cell
D002784 Cholesterol The principal sterol of all higher animals, distributed in body tissues, especially the brain and spinal cord, and in animal fats and oils. Epicholesterol
D004730 Endothelium, Vascular Single pavement layer of cells which line the luminal surface of the entire vascular system and regulate the transport of macromolecules and blood components. Capillary Endothelium,Vascular Endothelium,Capillary Endotheliums,Endothelium, Capillary,Endotheliums, Capillary,Endotheliums, Vascular,Vascular Endotheliums
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D000254 Sodium-Potassium-Exchanging ATPase An enzyme that catalyzes the active transport system of sodium and potassium ions across the cell wall. Sodium and potassium ions are closely coupled with membrane ATPase which undergoes phosphorylation and dephosphorylation, thereby providing energy for transport of these ions against concentration gradients. ATPase, Sodium, Potassium,Adenosinetriphosphatase, Sodium, Potassium,Na(+)-K(+)-Exchanging ATPase,Na(+)-K(+)-Transporting ATPase,Potassium Pump,Sodium Pump,Sodium, Potassium ATPase,Sodium, Potassium Adenosinetriphosphatase,Sodium-Potassium Pump,Adenosine Triphosphatase, Sodium, Potassium,Na(+) K(+)-Transporting ATPase,Sodium, Potassium Adenosine Triphosphatase,ATPase Sodium, Potassium,ATPase, Sodium-Potassium-Exchanging,Adenosinetriphosphatase Sodium, Potassium,Pump, Potassium,Pump, Sodium,Pump, Sodium-Potassium,Sodium Potassium Exchanging ATPase,Sodium Potassium Pump
D001665 Binding Sites The parts of a macromolecule that directly participate in its specific combination with another molecule. Combining Site,Binding Site,Combining Sites,Site, Binding,Site, Combining,Sites, Binding,Sites, Combining
D014471 Umbilical Veins Venous vessels in the umbilical cord. They carry oxygenated, nutrient-rich blood from the mother to the FETUS via the PLACENTA. In humans, there is normally one umbilical vein. Umbilical Vein,Vein, Umbilical,Veins, Umbilical

Related Publications

Y T Lau, and J K Chen, and M M Hsu, and H M Yu
May 1984, The American journal of physiology,
Y T Lau, and J K Chen, and M M Hsu, and H M Yu
January 1987, Basic research in cardiology,
Y T Lau, and J K Chen, and M M Hsu, and H M Yu
March 1994, Revista espanola de fisiologia,
Y T Lau, and J K Chen, and M M Hsu, and H M Yu
April 1978, British journal of pharmacology,
Y T Lau, and J K Chen, and M M Hsu, and H M Yu
October 1985, Klinische Wochenschrift,
Y T Lau, and J K Chen, and M M Hsu, and H M Yu
June 1981, Biochimica et biophysica acta,
Y T Lau, and J K Chen, and M M Hsu, and H M Yu
April 1988, Journal of neurochemistry,
Y T Lau, and J K Chen, and M M Hsu, and H M Yu
June 1983, British journal of clinical pharmacology,
Y T Lau, and J K Chen, and M M Hsu, and H M Yu
February 1985, Thrombosis and haemostasis,
Y T Lau, and J K Chen, and M M Hsu, and H M Yu
September 1986, The Journal of biological chemistry,
Copied contents to your clipboard!