Characterization of the redox components of transplasma membrane electron transport system from Leishmania donovani promastigotes

Biochim Biophys Acta. 2005 Oct 10;1725(3):314-26. doi: 10.1016/j.bbagen.2005.05.024. Epub 2005 Jun 22.

Abstract

An investigation has been made of the points of coupling of four nonpermeable electron acceptors e.g., alpha-lipoic acid (ALA), 5,5'-dithiobis (2-nitroaniline-N-sulphonic acid) (DTNS), 1,2-naphthoquinone-4-sulphonic acid (NQSA) and ferricyanide which are mainly reduced via an interaction with the redox sites present in the plasma membrane of Leishmania donovani promastigotes. ALA, DTNS, NQSA and ferricyanide reduction and part of O2 reduction is shown to take place on the exoplasmic face of the cell, for it is affected by external pH and agents that react with the external surface. Redox enzymes of the transplasma membrane electron transport system orderly transfer electron from one redox carrier to the next with the molecular oxygen as the final electron acceptor. The redox carriers mediate the transfer of electrons from metabolically generated reductant to nonpermeable electron acceptors and oxygen. At a pH of 6.4, respiration of Leishmania cells on glucose substrate shut down almost completely upon addition of an uncoupler FCCP and K+-ionophore valinomycin. The most pronounced effects on O2 uptake were obtained by treatment with antimycin A, 2-heptadecyl-4-hydroxyquinone-N-oxide, paracholoromercuribenzene sulphonic acid and trifluoperazine. Relatively smaller effects were obtained by treatment with potassium cyanide. Inhibition observed with respect to the reduction of the electron acceptors ALA, DTNS, NQSA and ferricyanide was not similar in most cases. The redox chain appears to be branched at several points and it is suggested that this redox chain incorporate iron-sulphur center, b-cytochromes, cyanide insensitive oxygen redox site, Na+ and K+ channel, capsaicin inhibited energy coupling site and trifluoperazine inhibited energy linked P-type ATPase. We analyzed the influence of ionic composition of the medium on reduction of electron acceptors in Leishmania donovani promastigotes. Our data suggest that K+ have some role for ALA reduction and Na+ for ferricyanide reduction. No significant effects were found with DTNS and NQSA reduction when Na+ or K+ was omitted from the medium. Stimulation of ALA, DTNS, NQSA and ferricyanide reduction was obtained by omitting Cl- from the medium. We propose that this redox system may be an energy source for control of membrane function in Leishmania cells.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aerobiosis / physiology
  • Amiloride / pharmacology
  • Anaerobiosis / physiology
  • Animals
  • Antimycin A / pharmacology
  • Cadmium Chloride / pharmacology
  • Capsaicin / pharmacology
  • Cell Membrane
  • Disulfides / metabolism
  • Electron Transport / drug effects*
  • Ferricyanides / metabolism
  • Hydrogen-Ion Concentration
  • Leishmania donovani / cytology
  • Leishmania donovani / drug effects
  • Leishmania donovani / metabolism*
  • Life Cycle Stages
  • Monensin / pharmacology
  • Naphthoquinones / metabolism
  • Oxidation-Reduction
  • Sulfanilic Acids / metabolism
  • Sulfhydryl Reagents / pharmacology
  • Sulfonic Acids / metabolism
  • Thioctic Acid / metabolism
  • Uncoupling Agents / pharmacology

Substances

  • 5,5'-dithiobis(2-nitroaniline-N-sulfonic acid)
  • Disulfides
  • Ferricyanides
  • Naphthoquinones
  • Sulfanilic Acids
  • Sulfhydryl Reagents
  • Sulfonic Acids
  • Uncoupling Agents
  • Antimycin A
  • Thioctic Acid
  • Amiloride
  • Monensin
  • Cadmium Chloride
  • 1,2-naphthoquinone-4-sulfonic acid
  • Capsaicin
  • potassium ferricyanide