Electrogenic nature of lysosomal proton pump as revealed with a cyanine dye

Shoji Ohkuma, Yoshinori Moriyama, Tatsuya Takano

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

Studies were carried out on the electrogenicity of the lysosomal proton pump using dipropyithiadicarbocyanine iodide (diS-C3-(5)) as a membrane potential probe. Pure lysosome preparations (tritosomes) quenched the fluorescence of diS-C3-(5). The quenching correlated well with the potassium ion diffusion potential (inside negative) generated by K+ with or without valinomycin. The quenching caused by lysosomes was reversed by lipophilic cations, tetraphenylarsonium (TPA) or triphenylmethyiphosphonium (TPMP). Mg-ATP also reversed the quenching, which was inhibited by a protonophore, 3,5-di-tert-butyl-4-hydroxybenzylidene-malononitrile (SF-6847). The properties of the ATP-induced recovery of the quenching were exactly the same as those of ATP-induced acidification, as measured with fluoresceinisothiocyanate-dextran (FD) (Ohkuma, S., et al. (1982) Proc. Natl. Acad. Sci. U.S. 79, 2758-2762) and acridine orange (Moriyama, Y., et al. (1982) J. Biochem. 92, 1333-1336), except replacement of the anion by an impermeable one enhanced ATPinduced recovery of quenching, but reduced ATP-induced acidification. Amines which dissipate △pH across the lysosomal membrane also enhanced the Mg-ATPinduced fluorescence recovery. These results suggest that isolated lysosomes exhibit an inside negative membrane potential, especially in low K+ medium, mostly due to the K+-diffusion potential, and that the Mg-ATP-driven proton pump causes membrane depolarization (in the direction of inside positive). These possibilities were supported by results on the uptake of the radioactive membrane-permeant ions [3H]TPMP and [14C]SCN. The present results provide evidence for the electrogenic nature of the lysosomal proton pump.

Original languageEnglish
Pages (from-to)1935-1943
Number of pages9
JournalJournal of Biochemistry
Volume94
Issue number6
Publication statusPublished - Dec 1983
Externally publishedYes

Fingerprint

Proton Pump
Proton Pumps
Adenosinetriphosphate
Quenching
Dyes
Protons
Coloring Agents
Adenosine Triphosphate
Pumps
Membranes
Lysosomes
Membrane Potential
Membrane
Recovery
Acidification
Membrane Potentials
Fluorescence
Enhanced recovery
Ions
Valinomycin

ASJC Scopus subject areas

  • Statistics, Probability and Uncertainty
  • Applied Mathematics
  • Physiology (medical)
  • Radiology Nuclear Medicine and imaging
  • Molecular Biology
  • Biochemistry

Cite this

Ohkuma, S., Moriyama, Y., & Takano, T. (1983). Electrogenic nature of lysosomal proton pump as revealed with a cyanine dye. Journal of Biochemistry, 94(6), 1935-1943.

Electrogenic nature of lysosomal proton pump as revealed with a cyanine dye. / Ohkuma, Shoji; Moriyama, Yoshinori; Takano, Tatsuya.

In: Journal of Biochemistry, Vol. 94, No. 6, 12.1983, p. 1935-1943.

Research output: Contribution to journalArticle

Ohkuma, S, Moriyama, Y & Takano, T 1983, 'Electrogenic nature of lysosomal proton pump as revealed with a cyanine dye', Journal of Biochemistry, vol. 94, no. 6, pp. 1935-1943.
Ohkuma, Shoji ; Moriyama, Yoshinori ; Takano, Tatsuya. / Electrogenic nature of lysosomal proton pump as revealed with a cyanine dye. In: Journal of Biochemistry. 1983 ; Vol. 94, No. 6. pp. 1935-1943.
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