Skip to main content
Biochemical Journal logoLink to Biochemical Journal
. 1981 Dec 15;200(3):555–563. doi: 10.1042/bj2000555

The effect of chloroquine on the metabolism of [35S]cystine in normal and cystinotic human skin fibroblasts.

C J Danpure
PMCID: PMC1163577  PMID: 7342970

Abstract

The present study concerns the effect of the lysosomotropic drug chloroquine on the uptake and metabolism of [35S]cystine in vitro by normal human fibroblasts and those from patients suffering from the lysosomal storage disease cystinosis. When the cells were cultured with [35S]cystine for periods in excess of 4 h, it was found that chloroquine considerably increased (up to 30-fold) the labelling of the intracellular cystine pool in cystinotic cells, with no increase or a much smaller increase in normal cells. For this effect chloroquine had an optimum concentration of 20 microM, with a small effect still being noticeable at 1 microM. A quinoline analogue, 4-(dimethylaminoethylamino)-7-iodoquinoline, had a similar effect to chloroquine. However, NH4Cl at concentrations of between 100 microM and 50 mM showed either no effect (at the lower concentrations) or a depression of intracellular cystine labelling (at the higher concentrations). The differences between the effects of the quinolines on cystinotic acid normal cells were not due to differences in total cell uptake of drug.

Full text

PDF
555

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Donato S. D., Wiesmann U. N., Herschkowitz N. Membrane adsorption and internalization of (14C)chloroquine by cultured human fibroblasts. Biochem Pharmacol. 1977 Jan 1;26(1):7–10. doi: 10.1016/0006-2952(77)90122-8. [DOI] [PubMed] [Google Scholar]
  2. Ehrenreich B. A., Cohn Z. A. The fate of peptides pinocytosed by macrophages in vitro. J Exp Med. 1969 Jan 1;129(1):227–245. doi: 10.1084/jem.129.1.227. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Gordon A. H., Hart P. D., Young M. R. Ammonia inhibits phagosome-lysosome fusion in macrophages. Nature. 1980 Jul 3;286(5768):79–80. doi: 10.1038/286079a0. [DOI] [PubMed] [Google Scholar]
  4. Griffiths P. A., Lloyd J. B. Evidence for lysosomal reduction of cystine residues. Biochem Biophys Res Commun. 1979 Jul 27;89(2):428–434. doi: 10.1016/0006-291x(79)90647-8. [DOI] [PubMed] [Google Scholar]
  5. Hart P. D., Young M. R. Manipulations of the phagosome-lysosome fusion response in cultured macrophages. Enhancement of fusion by chloroquine and other amines. Exp Cell Res. 1978 Jul;114(2):486–490. doi: 10.1016/0014-4827(78)90516-5. [DOI] [PubMed] [Google Scholar]
  6. Hart P. D., Young M. R. The effect of inhibitors and enhancers of phagosome--lysosome fusion in cultured macrophages on the phagosome membranes of ingested yeasts. Exp Cell Res. 1979 Feb;118(2):365–375. doi: 10.1016/0014-4827(79)90160-5. [DOI] [PubMed] [Google Scholar]
  7. Lie S. O., Schofield B. Inactivation of lysosomal function in normal cultured human fibroblasts by chloroquine. Biochem Pharmacol. 1973 Dec 1;22(23):3109–3114. doi: 10.1016/0006-2952(73)90197-4. [DOI] [PubMed] [Google Scholar]
  8. Mego J. L., Chung C. H. Effects of some antimalarials and related substances on intralysosomal proteolysis. Biochem Pharmacol. 1979;28(4):465–470. doi: 10.1016/0006-2952(79)90237-5. [DOI] [PubMed] [Google Scholar]
  9. Ohkuma S., Poole B. Fluorescence probe measurement of the intralysosomal pH in living cells and the perturbation of pH by various agents. Proc Natl Acad Sci U S A. 1978 Jul;75(7):3327–3331. doi: 10.1073/pnas.75.7.3327. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Oshima R. G., Rhead W. J., Thoene J. G., Schneider J. A. Cystine metabolism in human fibroblasts. Comparison of normal, cystinotic, and gamma-glutamylcysteine synethetase-deficient cells. J Biol Chem. 1976 Jul 25;251(14):4287–4293. [PubMed] [Google Scholar]
  11. Patrick A. D., Lake B. D. Cystinosis: electron microscopic evidence of lysosomal storage of cystine in lymph node. J Clin Pathol. 1968 Sep;21(5):571–575. doi: 10.1136/jcp.21.5.571. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Polet H. Chloroquine-3H: mechanism of uptake by Chang liver cells in vitro. J Pharmacol Exp Ther. 1976 Dec;199(3):687–694. [PubMed] [Google Scholar]
  13. Reijngoud D. J., Tager J. M. Chloroquine accumulation in isolated rat liver lysosomes. FEBS Lett. 1976 Apr 15;64(1):231–235. doi: 10.1016/0014-5793(76)80290-6. [DOI] [PubMed] [Google Scholar]
  14. Sando G. N., Titus-Dillon P., Hall C. W., Neufeld E. F. Inhibition of receptor-mediated uptake of a lysosomal enzyme into fibroblasts by chloroquine, procaine and ammonia. Exp Cell Res. 1979 Mar 15;119(2):359–364. doi: 10.1016/0014-4827(79)90364-1. [DOI] [PubMed] [Google Scholar]
  15. Schneider J. A., Rosenbloom F. M., Bradley K. H., Seegmiller J. E. Increased free-cystine content of fibroblasts cultured from patients with cystinosis. Biochem Biophys Res Commun. 1967 Nov 30;29(4):527–531. doi: 10.1016/0006-291x(67)90516-5. [DOI] [PubMed] [Google Scholar]
  16. Schulman J. D., Bradley K. H., Seegmiller J. E. Cystine: compartmentalization within lysosomes in cystinotic leukocytes. Science. 1969 Nov 28;166(3909):1152–1154. doi: 10.1126/science.166.3909.1152. [DOI] [PubMed] [Google Scholar]
  17. Schulman J. D., Bradley K. H. The metabolism of amino acids, peptides, and disulfides in lysosomes of fibroblasts cultured from normal individuals and those with cystinosis. J Exp Med. 1970 Dec 1;132(6):1090–1104. doi: 10.1084/jem.132.6.1090. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Schulman J. D., Schneider J. A., Bradley K. H., Seegmiller J. E. Cystine, cysteine, and glutathione metabolism in normal and cystinotic fibroblasts in vitro, and in cultured normal amniotic fluid cells. Clin Chim Acta. 1971 Dec;35(2):383–388. doi: 10.1016/0009-8981(71)90210-5. [DOI] [PubMed] [Google Scholar]
  19. Seglen P. O., Grinde B., Solheim A. E. Inhibition of the lysosomal pathway of protein degradation in isolated rat hepatocytes by ammonia, methylamine, chloroquine and leupeptin. Eur J Biochem. 1979 Apr 2;95(2):215–225. doi: 10.1111/j.1432-1033.1979.tb12956.x. [DOI] [PubMed] [Google Scholar]
  20. States B., Harris D., Segal S. Uptake and utilization of exogenous cystine by cystinotic and normal fibroblasts. J Clin Invest. 1974 Apr;53(4):1003–1016. doi: 10.1172/JCI107637. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Thoene J. G., Lemons R. Modulation of the intracellular cystine content of cystinotic fibroblasts by extracellular albumin. Pediatr Res. 1980 Jun;14(6):785–787. doi: 10.1203/00006450-198006000-00001. [DOI] [PubMed] [Google Scholar]
  22. Thoene J. G., Oshima R. G., Ritchie D. G., Schneider J. A. Cystinotic fibroblasts accumulate cystine from intracellular protein degradation. Proc Natl Acad Sci U S A. 1977 Oct;74(10):4505–4507. doi: 10.1073/pnas.74.10.4505. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Wibo M., Poole B. Protein degradation in cultured cells. II. The uptake of chloroquine by rat fibroblasts and the inhibition of cellular protein degradation and cathepsin B1. J Cell Biol. 1974 Nov;63(2 Pt 1):430–440. doi: 10.1083/jcb.63.2.430. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Wiesmann U. N., DiDonato S., Herschkowitz N. N. Effect of chloroquine on cultured fibroblasts: release of lysosomal hydrolases and inhibition of their uptake. Biochem Biophys Res Commun. 1975 Oct 27;66(4):1338–1343. doi: 10.1016/0006-291x(75)90506-9. [DOI] [PubMed] [Google Scholar]
  25. Willcox P., Rattray S. Secretion and uptake of beta-N-acetylglucosaminidase by fibroblasts. Effect of chloroquine and mannose 6-phosphate. Biochim Biophys Acta. 1979 Sep 3;586(3):442–452. doi: 10.1016/0304-4165(79)90034-5. [DOI] [PubMed] [Google Scholar]
  26. de Duve C., de Barsy T., Poole B., Trouet A., Tulkens P., Van Hoof F. Commentary. Lysosomotropic agents. Biochem Pharmacol. 1974 Sep 15;23(18):2495–2531. doi: 10.1016/0006-2952(74)90174-9. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES