Skip to main content
The Journal of Physiology logoLink to The Journal of Physiology
. 1980;300:19–30. doi: 10.1113/jphysiol.1980.sp013148

The effect of buffer composition and deoxygenation on the concentration of ionized magnesium inside human red blood cells.

P W Flatman
PMCID: PMC1279341  PMID: 6770081

Abstract

1. A method is described in which the concentration of ionized magnesium can be measured in intact red cells. The method uses an equilibrium dialysis technique originally developed by Ferreira & Lew (1976) and Flatman & Lew (1977) where the magnesium permeability of the red cell membrane is increased with the ionophore A23187. 2. The concentration of ionized magnesium in the oxygenated cells was found to be 0.39 mM and was not greatly affected by changes in the composition of the medium. 3. The concentration of ionized magnesium in deoxygenated cells showed more dependence on the composition of the medium. Values of 0.54 and 0.62 mM were found in cells incubated in Tris- and HCO3- buffered media respectively. The difference probably reflects increased competition between chloride and 2,3-diphosphoglycerate for common binding sites on haemoglobin in Tris-buffered cells. 3. Only a small increase of 0.16-0.22 mM was found in the concentration of ionized magnesium when the cells were deoxygenated. These changes are smaller than had been anticipated from estimates of the binding of ATP and 2,3-diphosphoglycerate to oxy- and deoxyhaemoglobin (Bunn, Ransil & Chao, 1971; Berger, Jänig, Gerber, Ruckpaul & Rapoport, 1973; Gerber, Berger, Jänig & Rapoport, 1973) and are unlikely to alter greatly the operation of magnesium-dependent metabolic or transport systems.

Full text

PDF
21

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Ashley C. C., Ellory J. C. The efflux of magnesium from single crustacean muscle fibres. J Physiol. 1972 Nov;226(3):653–674. doi: 10.1113/jphysiol.1972.sp010002. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Baker P. F., Crawford A. C. Mobility and transport of magnesium in squid giant axons. J Physiol. 1972 Dec;227(3):855–874. doi: 10.1113/jphysiol.1972.sp010062. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Benesch R. E., Benesch R., Yu C. I. The oxygenation of hemoglobin in the presence of 2,3-diphosphoglycerate. Effect of temperature, pH, ionic strength, and hemoglobin concentration. Biochemistry. 1969 Jun;8(6):2567–2571. doi: 10.1021/bi00834a046. [DOI] [PubMed] [Google Scholar]
  4. Berger H., Jänig G. R., Gerber G., Ruckpaul K., Rapoport S. M. Interaction of haemoglobin with ions. Interactions among magnesium, adenosine 5'-triphosphate, 2,3-bisphosphoglycerate, and oxygenated and deoxygenated human haemoglobin under simulated intracellular conditions. Eur J Biochem. 1973 Oct 18;38(3):553–562. doi: 10.1111/j.1432-1033.1973.tb03090.x. [DOI] [PubMed] [Google Scholar]
  5. Brinley F. J., Jr, Scarpa A. Ionized magnesium concentration in axoplasm of dialyzed squid axons. FEBS Lett. 1975 Jan 15;50(1):82–85. doi: 10.1016/0014-5793(75)81046-5. [DOI] [PubMed] [Google Scholar]
  6. Brinley F. J., Jr, Scarpa A., Tiffert T. The concentration of ionized magnesium in barnacle muscle fibres. J Physiol. 1977 Apr;266(3):545–565. doi: 10.1113/jphysiol.1977.sp011781. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Bunn H. F., Ransil B. J., Chao A. The interaction between erythrocyte organic phosphates, magnesium ion, and hemoglobin. J Biol Chem. 1971 Sep 10;246(17):5273–5279. [PubMed] [Google Scholar]
  8. Case G. D., Vanderkooi J. M., Scarpa A. Physical properties of biological membranes determined by the fluorescence of the calcium ionophore A23187. Arch Biochem Biophys. 1974 May;162(1):174–185. doi: 10.1016/0003-9861(74)90116-7. [DOI] [PubMed] [Google Scholar]
  9. Chiancone E., Norne J. E., Forsén S., Bonaventura J., Brunori M., Antonini E., Wyman J. Identification of chloride-binding sites in hemoglobin by nuclear-magnetic-resonance quadrupole-relaxation studies of hemoglobin digests. Eur J Biochem. 1975 Jul 1;55(2):385–390. doi: 10.1111/j.1432-1033.1975.tb02173.x. [DOI] [PubMed] [Google Scholar]
  10. De Weer P. Axoplasmic free magnesium levels and magnesium extrusion from squid giant axons. J Gen Physiol. 1976 Aug;68(2):159–178. doi: 10.1085/jgp.68.2.159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Ferreira H. G., Lew V. L. Use of ionophore A23187 to measure cytoplasmic Ca buffering and activation of the Ca pump by internal Ca. Nature. 1976 Jan 1;259(5538):47–49. doi: 10.1038/259047a0. [DOI] [PubMed] [Google Scholar]
  12. Flatman P. W., Lew V. L. The magnesium-dependence of sodium:potassium and sodium:sodium exchange mediated by the sodium pump in intact human red cells [proceedings]. J Physiol. 1979 Feb;287:33P–34P. [PubMed] [Google Scholar]
  13. Flatman P., Lew V. L. Use of ionophore A23187 to measure and to control free and bound cytoplasmic Mg in intact red cells. Nature. 1977 May 26;267(5609):360–362. doi: 10.1038/267360a0. [DOI] [PubMed] [Google Scholar]
  14. Gerber G., Berger H., Jänig G. R., Rapoport S. M. Interaction of haemoglobin with ions. Quantitative description of the state of magnesium, adenosine 5'-triphosphate, 2,3-bisphosphoglycerate, and human haemoglobin under simulated intracellular conditions. Eur J Biochem. 1973 Oct 18;38(3):563–571. doi: 10.1111/j.1432-1033.1973.tb03091.x. [DOI] [PubMed] [Google Scholar]
  15. Glynn I. M., Lew V. L., Lüthi U. Reversal of the potassium entry mechanism in red cells, with and without reversal of the entire pump cycle. J Physiol. 1970 Apr;207(2):371–391. doi: 10.1113/jphysiol.1970.sp009067. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Gupta R. K., Benovic J. L., Rose Z. B. The determination of the free magnesium level in the human red blood cell by 31P NMR. J Biol Chem. 1978 Sep 10;253(17):6172–6176. [PubMed] [Google Scholar]
  17. Kafka M. S., Holz R. W. Ionophores X537A and A23187. Effects on the permeability of lipid bimolecular membranes to dopamine and calcium. Biochim Biophys Acta. 1976 Feb 19;426(1):31–37. doi: 10.1016/0005-2736(76)90426-0. [DOI] [PubMed] [Google Scholar]
  18. Lew V. L., Ferreira H. G. Variable Ca sensitivity of a K-selective channel in intact red-cell membranes. Nature. 1976 Sep 23;263(5575):336–338. doi: 10.1038/263336a0. [DOI] [PubMed] [Google Scholar]
  19. ROGERS T. A. The exchange of radioactive magnesium in erythrocytes of several species. J Cell Comp Physiol. 1961 Apr;57:119–121. doi: 10.1002/jcp.1030570209. [DOI] [PubMed] [Google Scholar]
  20. Rubin H. Magnesium deprivation reproduces the coordinate effects of serum removal or cortisol addition on transport and metabolism in chick embryo fibroblasts. J Cell Physiol. 1976 Dec;89(4):613–625. doi: 10.1002/jcp.1040890418. [DOI] [PubMed] [Google Scholar]
  21. Veloso D., Guynn R. W., Oskarsson M., Veech R. L. The concentrations of free and bound magnesium in rat tissues. Relative constancy of free Mg 2+ concentrations. J Biol Chem. 1973 Jul 10;248(13):4811–4819. [PubMed] [Google Scholar]

Articles from The Journal of Physiology are provided here courtesy of The Physiological Society

RESOURCES