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. 1988 Mar;397:433–447. doi: 10.1113/jphysiol.1988.sp017010

Inhibition of chloride self-exchange with stilbene disulphonates in depolarized skeletal muscle of Rana temporaria.

J M Skydsgaard 1
PMCID: PMC1192134  PMID: 3261796

Abstract

1. The inhibition of 36Cl efflux with stilbene disulphonates, SD, has been studied under conditions of chloride equilibrium in depolarized fibre bundles from frog semitendinosi. The chosen probes were the aminoreactive derivative SITS and the derivative DNDS with no aminoreactive group. SD were added to the medium during 36Cl efflux allowing the estimation of fractional inhibition after a single 36Cl loading. 2. Both probes inhibited chloride self-exchange reversibly within the pH range 5.5-9.5 under study. 3. At SD concentrations above the half-inhibition concentration the inhibition reached a steady level with a time lag equal to that required for extracellular fluid change. The time constant for reversibility upon the removal of SD increased with decreasing pH, but rapid reversibility always appeared upon an increase of pH to 7.2. These findings suggest that SD may enter the membrane at low pH, but that the inhibitory action is confined to superficial membrane sites. 4. The inhibitory power of both probes showed a pronounced pH dependence, pK approximately 7. The half-inhibition concentration increased about 6-7 times when pH was lowered one unit from the pK value. 5. The apparent affinity of SITS to the transport system was about 5 times higher than that of DNDS. The apparent dissociation constants at neutral pH were 8.5 x 10(-5) M (SITS) and 4.5 x 10(-4) M (DNDS). Both probes showed a maximal inhibition close to 100% at neutral pH and approximately 85% at pH 5.5. 6. The inhibition depended on the chloride concentration in a way consistent with competitive inhibition in both neutral and acid media. 7. The results are consistent with the classical model of anion transport in frog muscle, suggesting that SD and chloride may compete for binding to a site with increasing anion affinity upon protonation; the results do not, however, exclude that the conductive and the non-conductive chloride transport modes in frog muscle are mediated by separate SD-sensitive transport pathways.

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Selected References

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  1. Barzilay M., Ship S., Cabantchik Z. I. Anion transport in red blood cells. I. Chemical properties of anion recognition sites as revealed by structure-activity relationships of aromatic sulfonic acids. Membr Biochem. 1979;2(2):227–254. doi: 10.3109/09687687909063866. [DOI] [PubMed] [Google Scholar]
  2. Bolton T. B., Vaughan-Jones R. D. Continuous direct measurement of intracellular chloride and pH in frog skeletal muscle. J Physiol. 1977 Sep;270(3):801–833. doi: 10.1113/jphysiol.1977.sp011983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cabantchik Z. I., Rothstein A. The nature of the membrane sites controlling anion permeability of human red blood cells as determined by studies with disulfonic stilbene derivatives. J Membr Biol. 1972 Dec 29;10(3):311–330. doi: 10.1007/BF01867863. [DOI] [PubMed] [Google Scholar]
  4. HODGKIN A. L., HOROWICZ P. The influence of potassium and chloride ions on the membrane potential of single muscle fibres. J Physiol. 1959 Oct;148:127–160. doi: 10.1113/jphysiol.1959.sp006278. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Hanke W., Miller C. Single chloride channels from Torpedo electroplax. Activation by protons. J Gen Physiol. 1983 Jul;82(1):25–45. doi: 10.1085/jgp.82.1.25. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hutter O. F., Warner A. E. Action of some foreign cations and anions on the chloride permeability of frog muscle. J Physiol. 1967 Apr;189(3):445–460. doi: 10.1113/jphysiol.1967.sp008178. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hutter O. F., Warner A. E. The effect of pH on the 36-Cl efflux from frog skeletal muscle. J Physiol. 1967 Apr;189(3):427–443. doi: 10.1113/jphysiol.1967.sp008177. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hutter O. F., Warner A. E. The pH sensitivity of the chloride conductance of frog skeletal muscle. J Physiol. 1967 Apr;189(3):403–425. doi: 10.1113/jphysiol.1967.sp008176. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hutter O. F., Warner A. E. The voltage dependence of the chloride conductance of frog muscle. J Physiol. 1972 Dec;227(1):275–290. doi: 10.1113/jphysiol.1972.sp010032. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kampmann L., Lepke S., Fasold H., Fritzsch G., Passow H. The kinetics of intramolecular cross-linking of the band 3 protein in the red blood cell membrane by 4,4'-diisothiocyano dihydrostilbene-2,2'-disulfonic acid (H2DIDS). J Membr Biol. 1982;70(3):199–216. doi: 10.1007/BF01870563. [DOI] [PubMed] [Google Scholar]
  11. Knauf P. A., Rothstein A. Chemical modification of membranes. I. Effects of sulfhydryl and amino reactive reagents on anion and cation permeability of the human red blood cell. J Gen Physiol. 1971 Aug;58(2):190–210. doi: 10.1085/jgp.58.2.190. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Loo D. D., McLarnon J. G., Vaughan P. C. Some observations on the behaviour of chloride current--voltage relations in Xenopus muscle membrane in acid solutions. Can J Physiol Pharmacol. 1981 Jan;59(1):7–13. doi: 10.1139/y81-002. [DOI] [PubMed] [Google Scholar]
  13. Moore L. E. Anion permeability of frog skeletal muscle. J Gen Physiol. 1969 Jul;54(1):33–52. doi: 10.1085/jgp.54.1.33. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Skydsgaard J. M. Influence of chloride concentration and pH on the 36Cl efflux from depolarized skeletal muscle of Rana temporaria. J Physiol. 1987 Apr;385:49–67. doi: 10.1113/jphysiol.1987.sp016483. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Vaughan P. C., McLarnon J. G., Loo D. D. Voltage dependence of chloride current through Xenopus muscle membrane in alkaline solutions. Can J Physiol Pharmacol. 1980 Sep;58(9):999–1010. doi: 10.1139/y80-153. [DOI] [PubMed] [Google Scholar]
  16. Vaughan P., Nok F. C. Effects of SITS on chloride permeation in Xenopus skeletal muscle. Can J Physiol Pharmacol. 1978 Dec;56(6):1051–1054. doi: 10.1139/y78-168. [DOI] [PubMed] [Google Scholar]
  17. Warner A. E. Kinetic properties of the chloride conductance of frog muscle. J Physiol. 1972 Dec;227(1):291–312. doi: 10.1113/jphysiol.1972.sp010033. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Woodbury J. W., Miles P. R. Anion conductance of frog muscle membranes: one channel, two kinds of pH dependence. J Gen Physiol. 1973 Sep;62(3):324–353. doi: 10.1085/jgp.62.3.324. [DOI] [PMC free article] [PubMed] [Google Scholar]

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