An estimate of sodium/potassium pump activity and the number of pump sites in the smooth muscle of the guinea pig taenia coli, using [3H]ouabain

A. F. Brading, Jonathan Widdicombe

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Abstract

Binding of tritiated ouabain to the smooth muscle of the guinea pig taenia coli showed two components, one saturable at lower glycoside concentrations and the other linear with increasing concentrations. The saturable component alone was affected by extracellular potassium concentrations. This component seems to be bound to sodium pumping sites, and when completely saturated binds 1.1 x 1011 molecules per mg fresh wt. of tissue, or 250-300 molecules per square micron of membrane, assuming a volume:surface ratio of 1.5 μm. Only a fraction of 42K uptake by the cells can be blocked by ouabain at maximal concentrations. In normal Krebs solutions two thirds can be blocked The remaining one third is presumably passive uptake. The fraction blocked is reduced as the extracellular postassium concentration, and thus passive uptake is increased. The amount of potassium pumped into the cells at various concentrations of extracellular potassium has been calculated. In normal Krebs solution the amount pumped in 45 min was 20.0 mmole/kg fresh wt., and this was increased at higher potassium concentrations. On the assumption of a stoichiometry of 3Na:2K, the pump sites in normal Krebs solution have a turnover rate of 1320 min-1. Indirect calculations of sodium movements suggest that the sodium permeability may be about 0.9 x 10-8 cm sec-1 and the pump may generate a current of 30-60 kΩcm2. This crossing an Ohmic membrane resistance of 30-60 kΩcm2 would be equivalent to a potential difference of 3-5 mV.

Original languageEnglish (US)
Pages (from-to)235-249
Number of pages15
JournalJournal of Physiology
Volume238
Issue number2
StatePublished - 1974
Externally publishedYes

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Sodium-Potassium-Exchanging ATPase
Ouabain
Smooth Muscle
Potassium
Guinea Pigs
Colon
Sodium
Membranes
Glycosides
Electric Impedance
Permeability
Krebs-Ringer solution

ASJC Scopus subject areas

  • Physiology

Cite this

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title = "An estimate of sodium/potassium pump activity and the number of pump sites in the smooth muscle of the guinea pig taenia coli, using [3H]ouabain",
abstract = "Binding of tritiated ouabain to the smooth muscle of the guinea pig taenia coli showed two components, one saturable at lower glycoside concentrations and the other linear with increasing concentrations. The saturable component alone was affected by extracellular potassium concentrations. This component seems to be bound to sodium pumping sites, and when completely saturated binds 1.1 x 1011 molecules per mg fresh wt. of tissue, or 250-300 molecules per square micron of membrane, assuming a volume:surface ratio of 1.5 μm. Only a fraction of 42K uptake by the cells can be blocked by ouabain at maximal concentrations. In normal Krebs solutions two thirds can be blocked The remaining one third is presumably passive uptake. The fraction blocked is reduced as the extracellular postassium concentration, and thus passive uptake is increased. The amount of potassium pumped into the cells at various concentrations of extracellular potassium has been calculated. In normal Krebs solution the amount pumped in 45 min was 20.0 mmole/kg fresh wt., and this was increased at higher potassium concentrations. On the assumption of a stoichiometry of 3Na:2K, the pump sites in normal Krebs solution have a turnover rate of 1320 min-1. Indirect calculations of sodium movements suggest that the sodium permeability may be about 0.9 x 10-8 cm sec-1 and the pump may generate a current of 30-60 kΩcm2. This crossing an Ohmic membrane resistance of 30-60 kΩcm2 would be equivalent to a potential difference of 3-5 mV.",
author = "Brading, {A. F.} and Jonathan Widdicombe",
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T1 - An estimate of sodium/potassium pump activity and the number of pump sites in the smooth muscle of the guinea pig taenia coli, using [3H]ouabain

AU - Brading, A. F.

AU - Widdicombe, Jonathan

PY - 1974

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N2 - Binding of tritiated ouabain to the smooth muscle of the guinea pig taenia coli showed two components, one saturable at lower glycoside concentrations and the other linear with increasing concentrations. The saturable component alone was affected by extracellular potassium concentrations. This component seems to be bound to sodium pumping sites, and when completely saturated binds 1.1 x 1011 molecules per mg fresh wt. of tissue, or 250-300 molecules per square micron of membrane, assuming a volume:surface ratio of 1.5 μm. Only a fraction of 42K uptake by the cells can be blocked by ouabain at maximal concentrations. In normal Krebs solutions two thirds can be blocked The remaining one third is presumably passive uptake. The fraction blocked is reduced as the extracellular postassium concentration, and thus passive uptake is increased. The amount of potassium pumped into the cells at various concentrations of extracellular potassium has been calculated. In normal Krebs solution the amount pumped in 45 min was 20.0 mmole/kg fresh wt., and this was increased at higher potassium concentrations. On the assumption of a stoichiometry of 3Na:2K, the pump sites in normal Krebs solution have a turnover rate of 1320 min-1. Indirect calculations of sodium movements suggest that the sodium permeability may be about 0.9 x 10-8 cm sec-1 and the pump may generate a current of 30-60 kΩcm2. This crossing an Ohmic membrane resistance of 30-60 kΩcm2 would be equivalent to a potential difference of 3-5 mV.

AB - Binding of tritiated ouabain to the smooth muscle of the guinea pig taenia coli showed two components, one saturable at lower glycoside concentrations and the other linear with increasing concentrations. The saturable component alone was affected by extracellular potassium concentrations. This component seems to be bound to sodium pumping sites, and when completely saturated binds 1.1 x 1011 molecules per mg fresh wt. of tissue, or 250-300 molecules per square micron of membrane, assuming a volume:surface ratio of 1.5 μm. Only a fraction of 42K uptake by the cells can be blocked by ouabain at maximal concentrations. In normal Krebs solutions two thirds can be blocked The remaining one third is presumably passive uptake. The fraction blocked is reduced as the extracellular postassium concentration, and thus passive uptake is increased. The amount of potassium pumped into the cells at various concentrations of extracellular potassium has been calculated. In normal Krebs solution the amount pumped in 45 min was 20.0 mmole/kg fresh wt., and this was increased at higher potassium concentrations. On the assumption of a stoichiometry of 3Na:2K, the pump sites in normal Krebs solution have a turnover rate of 1320 min-1. Indirect calculations of sodium movements suggest that the sodium permeability may be about 0.9 x 10-8 cm sec-1 and the pump may generate a current of 30-60 kΩcm2. This crossing an Ohmic membrane resistance of 30-60 kΩcm2 would be equivalent to a potential difference of 3-5 mV.

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