Role of calcium and calmodulin in the regulation of the rabbit ileal brush-border membrane Na+/H+ antiporter

Eugene Emmer, Richard P. Rood, John H. Wesolek, Michael E. Cohen, R. Scott Braithwaite, Geoffrey W.G. Sharp, Heini Murer, Mark Donowitz

Research output: Contribution to journalArticle

Abstract

In rabbit ileum, Ca2+/calmodulin (CaM) appears to be involved in physiologically inhibiting the linked NaCl absorptive process, since inhibitors of Ca2+/CaM stimulate linked Na+ and Cl- absorption. The role of Ca2+/CaM-dependent phosphorylation in regulation of the brush-border Na+/H+ antiporter, which is believed to be part of the neutral linked NaCl absorptive process, was studied using purified brush-border membrane vesicles, which contain both the Na+/H+ antiporter and Ca2+/CaM-dependent protein kinase(s) and its phosphoprotein substrates. Rabbit ileal villus cell brush-border membrane vesicles were prepared by Mg precipitation and depleted of ATP. Using a freezethaw technique, the ATP-depleted vesicles were loaded with Ca2+, CaM, ATP and an ATP-regenerating system consisting of creatine kinase and creatine phosphate. The combination of Ca2+/CaM and ATP inhibited Na+/H+ exchange by 45±13%. This effect was specific since Ca2+/CaM and ATP did not alter diffusive Na+ uptake, Na+-dependent glucose entry, or Na+ or glucose equilibrium volumes. The inhibition of the Na+/H+ exchanger by Ca2+/CaM/ATP was due to an effect on the Vmax and not on the Km for Na+. In the presence of CaM and ATP, Ca2+ caused a concentration-dependent inhibition of Na+ uptake, with an effect 50% of maximum occurring at 120 nm. This Ca2+ concentration dependence was similar to the Ca2+ concentration dependence of Ca2+/CaM-dependent phosphorylation of specific proteins in the vesicles. The Ca2+/CaM/ATP-inhibition of Na+/H+ exchange was reversed by W13, a Ca2+/CaM antagonist, but not by a hydrophobic control, W12, or by H-7, a protein kinase C antagonist. we conclude that Ca2+, acting through CaM, regulates ileal brush-border Na+/H+ exchange, and that this may be involved in the regulation of neutral linked NaCl absorption.

Original languageEnglish (US)
Pages (from-to)207-215
Number of pages9
JournalThe Journal of Membrane Biology
Volume108
Issue number3
DOIs
StatePublished - Jun 1 1989

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Keywords

  • absorption
  • brush-border vesicle
  • calcium
  • calmodulin
  • ileum
  • phosphorylation

ASJC Scopus subject areas

  • Biophysics
  • Physiology
  • Cell Biology

Cite this

Emmer, E., Rood, R. P., Wesolek, J. H., Cohen, M. E., Braithwaite, R. S., Sharp, G. W. G., Murer, H., & Donowitz, M. (1989). Role of calcium and calmodulin in the regulation of the rabbit ileal brush-border membrane Na+/H+ antiporter. The Journal of Membrane Biology, 108(3), 207-215. https://doi.org/10.1007/BF01871735