On the number of sodium pumping sites in cell membranes

Baker, P. F.; Willis, John S.

doi:10.1016/0005-2736(69)90180-1

Cited by 65 publications

(29 citation statements)

References 5 publications

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“…After incubation A. e H)·ouabain (4.5 x lQ6cprrv'fl ask) has heen added to the standard medium , giving final ouabain-concentratiOTli of 9 x 10-' and IO-l M respectively. Ccll-bound e H)·ouabain has been de termined after 4 hr of e H)+ouabain intubation al 37".…”

Section: Specijic E H )-Ouabain Bindingmentioning

confidence: 99%

Cardiac glycoside receptors in cultured heart cells—I. characterization of one single class of high affinity receptors in heart muscle cells from chick embryos

Werdan

Wagenknecht

Zwißler

et al. 1984

Biochemical Pharmacology

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Section: Specijic E H )-Ouabain Bindingmentioning

confidence: 99%

Cardiac glycoside receptors in cultured heart cells—I. characterization of one single class of high affinity receptors in heart muscle cells from chick embryos

Werdan

Wagenknecht

Zwißler

et al. 1984

Biochemical Pharmacology

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“…One reason for the obstruction to use of cultured cells is the difficulty of controlling the intracellular concentrations of alkali metal cations, which seems partly to be due to uneven distribution of the cations inside the cells (PALMER and CIVAN, 1977;GUPTA and HALL, 1978). HeLa cells are known to have much denser [3H] ouabain-binding sites on the cell membrane than red cells (BAKER and WILLIS, 1969). This seemed partly to overcome the disadvantage of using cultured cells in the study of cation transport.…”

Section: Cellsmentioning

confidence: 99%

Interrelation between membrane transport and the contents of alkali metal cations in HeLa cells.

et al. 1982

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“…7B illustrates two further pieces of evidence for a reciprocal relationship between activation of the Na pump and inhibition of the saturable component of glycoside binding. These are (1) that partial replacement of external Na by choline, which increases the apparent affinity of the Na pump for external K ions (Whittam & Ager, 1964;Baker & Connelly, 1966;Garrahan & Glynn, 1967;Baker et al 1969), also decreases the K concentration required to reduce the rate of glycoside binding to one half, and (2) that Rb and Cs ions, which can replace K as activators of the Na pump (Baker & Connelly, 1966;Sjodin & Beauge, 1967;Baker et al 1969) also reduce the rate of glycoside binding. In both processes Rb is an excellent substitute for K whereas Cs is about one tenth as effective.…”

Section: Ouabain Binding To Intact Cells 451mentioning

confidence: 99%

Binding of the cardiac glycoside ouabain to intact cells

Baker¹,

Willis²

1972

The Journal of Physiology

199

116

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SUMMARY1. Measurements were made of the binding of 73H]ouabain to a variety of cell types.2. Two components of binding could usually be distinguished: a component that saturated at low glycoside concentrations and a component that increased up to the highest ouabain concentrations examined.3. Detailed studies with HeLa cells and kidney slices from guinea-pigs showed that the saturable component is probably associated with inhibition of the Na pump. The main evidence for this is (a) at low concentrations of ouabain there is a close correspondence between the concentration of ouabain giving half-maximum binding and the concentration giving halfmaximum inhibition of the Na pump; (b) at low glycoside concentrations, binding precedes inhibition of the Na pump; (c) the rate of binding is very sensitive to external K ions, being highest in the absence of K; (d) binding is reversible and the release of ouabain is associated with reactivation of the Na pump, (e) binding is reduced in the absence of Na ions and in the presence of metabolic inhibitors; (f) binding has a Q10 of about 4; and (g) in the presence of Na and ATP, lysed HeLa cells bind a similar amount of ouabain and the binding is sensitive to K ions.4. The linear component of binding does not seem to involve the Na pump and it may reflect uptake of ouabain into the cell interior. It has a Q10 of 2-5 and is unaffected by K concentrations which have a large effect on the saturable component.5. Bound ouabain could be removed from HeLa cells by low pH, trichloroacetic acid, urea, high temperatures and 100 % ethanol. These agents did not distinguish between the two components of binding. 6. Criteria are developed for estimating the number of Na pumping sites in cells and the data for ouabain-binding to a number of cells is compared * Present address:

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On the number of sodium pumping sites in cell membranes

Cited by 65 publications

References 5 publications

Cardiac glycoside receptors in cultured heart cells—I. characterization of one single class of high affinity receptors in heart muscle cells from chick embryos

Cardiac glycoside receptors in cultured heart cells—I. characterization of one single class of high affinity receptors in heart muscle cells from chick embryos

Interrelation between membrane transport and the contents of alkali metal cations in HeLa cells.

Binding of the cardiac glycoside ouabain to intact cells

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