Phorbol-ester is bound specifically to renal luminal membranes and stimulates the Na-H antiporter

Z. Talor, George Mejicano

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

We characterized binding of [3H]-phorbol-12,13-dibutyrate (an activator of protein kinase C) to highly purified rabbit cortical renal luminal membranes and measured its effect on the kinetics of the Na-H antiporter. There was 95% specific binding to luminal membranes and this binding was time, temperature and pH dependent with optimal binding at 4°C and pH 7.4. Scatchard analysis of the binding revealed Kd of 0.8 μM and Bmax of 19.4 pmoles/mg protein. Phorbol-12,13-dibutyrate stimulated the Vmax of the Na-H antiporter by 16.5% ± 4.7 at 10-6 M and by 19.9% ± 5.4 at 10-5 M. The protein kinase C inhibitor H71 prevented the observed stimulation. Thus, there is a correlation between phorbol ester binding and phorbol ester stimulation of the Na-H antiporter. These data demonstrate that protein kinase C plays a role in the stimulation of the Na-H antiporter in renal luminal membranes.

Original languageEnglish (US)
Pages (from-to)335-343
Number of pages9
JournalResearch Communications in Chemical Pathology and Pharmacology
Volume61
Issue number3
StatePublished - 1988
Externally publishedYes

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Sodium-Hydrogen Antiporter
Phorbol Esters
Protein Kinase C
Phorbol 12,13-Dibutyrate
Membranes
Kidney
Protein C Inhibitor
Protein Kinase Inhibitors
Rabbits
Kinetics
Temperature
Proteins

ASJC Scopus subject areas

  • Pharmacology
  • Toxicology

Cite this

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abstract = "We characterized binding of [3H]-phorbol-12,13-dibutyrate (an activator of protein kinase C) to highly purified rabbit cortical renal luminal membranes and measured its effect on the kinetics of the Na-H antiporter. There was 95{\%} specific binding to luminal membranes and this binding was time, temperature and pH dependent with optimal binding at 4°C and pH 7.4. Scatchard analysis of the binding revealed Kd of 0.8 μM and Bmax of 19.4 pmoles/mg protein. Phorbol-12,13-dibutyrate stimulated the Vmax of the Na-H antiporter by 16.5{\%} ± 4.7 at 10-6 M and by 19.9{\%} ± 5.4 at 10-5 M. The protein kinase C inhibitor H71 prevented the observed stimulation. Thus, there is a correlation between phorbol ester binding and phorbol ester stimulation of the Na-H antiporter. These data demonstrate that protein kinase C plays a role in the stimulation of the Na-H antiporter in renal luminal membranes.",
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T1 - Phorbol-ester is bound specifically to renal luminal membranes and stimulates the Na-H antiporter

AU - Talor, Z.

AU - Mejicano, George

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N2 - We characterized binding of [3H]-phorbol-12,13-dibutyrate (an activator of protein kinase C) to highly purified rabbit cortical renal luminal membranes and measured its effect on the kinetics of the Na-H antiporter. There was 95% specific binding to luminal membranes and this binding was time, temperature and pH dependent with optimal binding at 4°C and pH 7.4. Scatchard analysis of the binding revealed Kd of 0.8 μM and Bmax of 19.4 pmoles/mg protein. Phorbol-12,13-dibutyrate stimulated the Vmax of the Na-H antiporter by 16.5% ± 4.7 at 10-6 M and by 19.9% ± 5.4 at 10-5 M. The protein kinase C inhibitor H71 prevented the observed stimulation. Thus, there is a correlation between phorbol ester binding and phorbol ester stimulation of the Na-H antiporter. These data demonstrate that protein kinase C plays a role in the stimulation of the Na-H antiporter in renal luminal membranes.

AB - We characterized binding of [3H]-phorbol-12,13-dibutyrate (an activator of protein kinase C) to highly purified rabbit cortical renal luminal membranes and measured its effect on the kinetics of the Na-H antiporter. There was 95% specific binding to luminal membranes and this binding was time, temperature and pH dependent with optimal binding at 4°C and pH 7.4. Scatchard analysis of the binding revealed Kd of 0.8 μM and Bmax of 19.4 pmoles/mg protein. Phorbol-12,13-dibutyrate stimulated the Vmax of the Na-H antiporter by 16.5% ± 4.7 at 10-6 M and by 19.9% ± 5.4 at 10-5 M. The protein kinase C inhibitor H71 prevented the observed stimulation. Thus, there is a correlation between phorbol ester binding and phorbol ester stimulation of the Na-H antiporter. These data demonstrate that protein kinase C plays a role in the stimulation of the Na-H antiporter in renal luminal membranes.

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