Skip to main content
PMC home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1986 Mar 1;102(3):967–971. doi: 10.1083/jcb.102.3.967

pH-regulated anion antiport in nucleated mammalian cells

PMCID: PMC2114147  PMID: 3949885

Abstract

The uptake of 36Cl- into cells was measured after preincubation in medium containing nigericin and KCl to allow control of the intracellular pH. When the pH was increased from pH 7.0 to pH 7.3 there was a 10-fold increase in the rate of 36Cl- uptake. The increase was half maximal at pH 7.15 in Vero and L-cells, whereas in phorbol 12- myristate 13-acetate-treated Vero cells the increase was half maximal at pH 6.9. Kinetic studies showed that in cells preincubated with nigericin and isotonic KCl, both at pH 7.0 and at pH 8.0, the Km for Cl- was 7 mM. In the two cases the Jmax was 1.7 X 10(8) Cl- ions X cell-1 X s-1 and 1.6 X 10(9) Cl- ions X cell-1 X s-1, respectively. Bicarbonate inhibited 36Cl- uptake with a Ki of 5-6 mM. Probably, the anion antiporter plays a role in the regulation of the intracellular pH.

Full Text

The Full Text of this article is available as a PDF (541.3 KB).

Selected References

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

  1. Aickin C. C., Thomas R. C. An investigation of the ionic mechanism of intracellular pH regulation in mouse soleus muscle fibres. J Physiol. 1977 Dec;273(1):295–316. doi: 10.1113/jphysiol.1977.sp012095. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Besterman J. M., Cuatrecasas P. Phorbol esters rapidly stimulate amiloride-sensitive Na+/H+ exchange in a human leukemic cell line. J Cell Biol. 1984 Jul;99(1 Pt 1):340–343. doi: 10.1083/jcb.99.1.340. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Boron W. F. Transport of H+ and of ionic weak acids and bases. J Membr Biol. 1983;72(1-2):1–16. doi: 10.1007/BF01870311. [DOI] [PubMed] [Google Scholar]
  4. Brahm J. Temperature-dependent changes of chloride transport kinetics in human red cells. J Gen Physiol. 1977 Sep;70(3):283–306. doi: 10.1085/jgp.70.3.283. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cabantchik Z. I., Knauf P. A., Rothstein A. The anion transport system of the red blood cell. The role of membrane protein evaluated by the use of 'probes'. Biochim Biophys Acta. 1978 Sep 29;515(3):239–302. doi: 10.1016/0304-4157(78)90016-3. [DOI] [PubMed] [Google Scholar]
  6. Frelin C., Vigne P., Lazdunski M. The amiloride-sensitive Na+/H+ antiport in 3T3 fibroblasts. J Biol Chem. 1983 May 25;258(10):6272–6276. [PubMed] [Google Scholar]
  7. Grinstein S., Cohen S., Goetz J. D., Rothstein A., Gelfand E. W. Characterization of the activation of Na+/H+ exchange in lymphocytes by phorbol esters: change in cytoplasmic pH dependence of the antiport. Proc Natl Acad Sci U S A. 1985 Mar;82(5):1429–1433. doi: 10.1073/pnas.82.5.1429. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Grinstein S., Goetz J. D., Rothstein A. 22Na+ fluxes in thymic lymphocytes. II. Amiloride-sensitive Na+/H+ exchange pathway; reversibility of transport and asymmetry of the modifier site. J Gen Physiol. 1984 Oct;84(4):585–600. doi: 10.1085/jgp.84.4.585. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Jentsch T. J., Stahlknecht T. R., Hollwede H., Fischer D. G., Keller S. K., Wiederholt M. A bicarbonate-dependent process inhibitable by disulfonic stilbenes and a Na+/H+ exchange mediate 22Na+ uptake into cultured bovine corneal endothelium. J Biol Chem. 1985 Jan 25;260(2):795–801. [PubMed] [Google Scholar]
  10. L'Allemain G., Paris S., Pouysségur J. Role of a Na+-dependent Cl-/HCO3- exchange in regulation of intracellular pH in fibroblasts. J Biol Chem. 1985 Apr 25;260(8):4877–4883. [PubMed] [Google Scholar]
  11. Larkin J. M., Donzell W. C., Anderson R. G. Modulation of intracellular potassium and ATP: effects on coated pit function in fibroblasts and hepatocytes. J Cell Physiol. 1985 Sep;124(3):372–378. doi: 10.1002/jcp.1041240303. [DOI] [PubMed] [Google Scholar]
  12. Mekada E., Kohno K., Ishiura M., Uchida T., Okada Y. Methylamine facilitates demonstration of specific uptake of diphtheria toxin by CHO cell and toxin-resistant CHO cell mutants. Biochem Biophys Res Commun. 1982 Dec 15;109(3):792–799. doi: 10.1016/0006-291x(82)92009-5. [DOI] [PubMed] [Google Scholar]
  13. Middlebrook J. L., Dorland R. B., Leppla S. H. Association of diphtheria toxin with Vero cells. Demonstration of a receptor. J Biol Chem. 1978 Oct 25;253(20):7325–7330. [PubMed] [Google Scholar]
  14. Moolenaar W. H., Tertoolen L. G., de Laat S. W. Phorbol ester and diacylglycerol mimic growth factors in raising cytoplasmic pH. Nature. 1984 Nov 22;312(5992):371–374. doi: 10.1038/312371a0. [DOI] [PubMed] [Google Scholar]
  15. Moolenaar W. H., Tertoolen L. G., de Laat S. W. The regulation of cytoplasmic pH in human fibroblasts. J Biol Chem. 1984 Jun 25;259(12):7563–7569. [PubMed] [Google Scholar]
  16. Moolenaar W. H., Tsien R. Y., van der Saag P. T., de Laat S. W. Na+/H+ exchange and cytoplasmic pH in the action of growth factors in human fibroblasts. Nature. 1983 Aug 18;304(5927):645–648. doi: 10.1038/304645a0. [DOI] [PubMed] [Google Scholar]
  17. Olsnes S., Sandvig K., Madshus I. H., Sundan A. Entry mechanisms of protein toxins and picornaviruses. Biochem Soc Symp. 1985;50:171–191. [PubMed] [Google Scholar]
  18. Paradiso A. M., Tsien R. Y., Machen T. E. Na+-H+ exchange in gastric glands as measured with a cytoplasmic-trapped, fluorescent pH indicator. Proc Natl Acad Sci U S A. 1984 Dec;81(23):7436–7440. doi: 10.1073/pnas.81.23.7436. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Paris S., Pouysségur J. Biochemical characterization of the amiloride-sensitive Na+/H+ antiport in Chinese hamster lung fibroblasts. J Biol Chem. 1983 Mar 25;258(6):3503–3508. [PubMed] [Google Scholar]
  20. Paris S., Pouysségur J. Growth factors activate the Na+/H+ antiporter in quiescent fibroblasts by increasing its affinity for intracellular H+. J Biol Chem. 1984 Sep 10;259(17):10989–10994. [PubMed] [Google Scholar]
  21. Roos A., Boron W. F. Intracellular pH. Physiol Rev. 1981 Apr;61(2):296–434. doi: 10.1152/physrev.1981.61.2.296. [DOI] [PubMed] [Google Scholar]
  22. Rosoff P. M., Stein L. F., Cantley L. C. Phorbol esters induce differentiation in a pre-B-lymphocyte cell line by enhancing Na+/H+ exchange. J Biol Chem. 1984 Jun 10;259(11):7056–7060. [PubMed] [Google Scholar]
  23. Rothenberg P., Glaser L., Schlesinger P., Cassel D. Activation of Na+/H+ exchange by epidermal growth factor elevates intracellular pH in A431 cells. J Biol Chem. 1983 Oct 25;258(20):12644–12653. [PubMed] [Google Scholar]
  24. Rothstein A., Cabantchik Z. I., Knauf P. Mechanism of anion transport in red blood cells: role of membrane proteins. Fed Proc. 1976 Jan;35(1):3–10. [PubMed] [Google Scholar]
  25. Sandvig K., Olsnes S. Anion requirement and effect of anion transport inhibitors on the response of vero cells to diphtheria toxin and modeccin. J Cell Physiol. 1984 Apr;119(1):7–14. doi: 10.1002/jcp.1041190103. [DOI] [PubMed] [Google Scholar]
  26. Swann K., Whitaker M. Stimulation of the Na/H exchanger of sea urchin eggs by phorbol ester. Nature. 1985 Mar 21;314(6008):274–277. doi: 10.1038/314274a0. [DOI] [PubMed] [Google Scholar]
  27. Vigne P., Frelin C., Lazdunski M. The Na+-dependent regulation of the internal pH in chick skeletal muscle cells. The role of the Na+/H+ exchange system and its dependence on internal pH. EMBO J. 1984 Aug;3(8):1865–1870. doi: 10.1002/j.1460-2075.1984.tb02060.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES