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Biophysical Journal 54: 241-247 (1988)
© 1988 the Biophysical Society

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Hypophosphite ion as a 31P nuclear magnetic resonance probe of membrane potential in erythrocyte suspensions.

K Kirk, P W Kuchel and R J Labotka

Department of Biochemistry, University of Sydney, New South Wales, Australia.

ABSTRACT

Hypophosphorus acid has a single pKa of 1.1 and at physiological pH values it is therefore present almost entirely as the univalent hypophosphite ion. When added to a red cell suspension the ion crosses the cell membrane rapidly, via the anion exchange protein, and the intra- and extracellular populations of the ion give rise to separate 31P NMR resonances. From a single 31P NMR spectrum it was possible to determine the relative amounts of hypophosphite in the intra- and extracellular compartments and thereby estimate the corresponding concentrations. The ratio of intracellular to extracellular hypophosphite concentration was independent of the total hypophosphite concentration for cells suspended in NaCl solutions and was independent of hematocrit. The hypophosphite distribution ratio increased as extracellular NaCl was replaced iso-osmotically with citrate or sucrose, through it remained very similar to the corresponding hydrogen ion distribution ratio. Incorporation of the hypophosphite distribution ratio into the Nernst equation yielded an estimate of the membrane potential. For cells suspended in NaCl solutions the estimated potential was consistently around -10 mV.




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T. J. Larkin, W. A. Bubb, and P. W. Kuchel
pH and Cell Volume Effects on H2O and Phosphoryl Resonance Splitting in Rapid-Spinning NMR of Red Cells
Biophys. J., March 1, 2007; 92(5): 1770 - 1776.
[Abstract] [Full Text] [PDF]


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Copyright © 1988 by the Biophysical Society.