Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1982 Jul;79(14):4442–4445. doi: 10.1073/pnas.79.14.4442

Cystine accumulation and loss in normal, heterozygous, and cystinotic fibroblasts.

A J Jonas, A A Greene, M L Smith, J A Schneider
PMCID: PMC346688  PMID: 6956872

Abstract

Cystinotic fibroblasts contain approximately 100 times more cystine than do normal control fibroblasts. When cystinotic fibroblasts were placed in the presence of 30 mM cysteine-glutathione mixed disulfide (CSSG) for 24 hr, their cystine content increased about 3-fold. Similar treatment of normal fibroblasts and fibroblasts from patients heterozygous for cystinosis resulted in a 6- to 7-fold increase in cystine content. In all three cell types, the intracellular free cystine is located within lysosomes. When placed in cystine-free medium after 24 hr in CSSG-containing medium, the normal and heterozygous fibroblasts rapidly lost their lysosomal cystine (t 1/2 = 20 min), but the cystine content of the cystinotic cells remained stable for over 90 min. In contrast to the findings in intact fibroblasts, cystine loss could not be demonstrated from isolated, cystine-loaded lysosomes from any of the three cell types.

Full text

PDF
4442

Selected References

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

  1. Baccino F. M., Zuretti M. F. Structural equivalents of latency for lysosome hydrolases. Biochem J. 1975 Jan;146(1):97–108. doi: 10.1042/bj1460097. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Coon H. G., Weiss M. C. A quantitative comparison of formation of spontaneous and virus-produced viable hybrids. Proc Natl Acad Sci U S A. 1969 Mar;62(3):852–859. doi: 10.1073/pnas.62.3.852. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. DULBECCO R., VOGT M. Plaque formation and isolation of pure lines with poliomyelitis viruses. J Exp Med. 1954 Feb;99(2):167–182. doi: 10.1084/jem.99.2.167. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Eriksson B., Eriksson S. A. Synthesis and characterization of the L-cysteine-glutathione mixed disulfide. Acta Chem Scand. 1967;21(5):1304–1312. doi: 10.3891/acta.chem.scand.21-1304. [DOI] [PubMed] [Google Scholar]
  5. Fahey R. C., Brody S., Mikolajczyk S. D. Changes in the glutathione thiol-disulfide status of Neurospora crassa conidia during germination and aging. J Bacteriol. 1975 Jan;121(1):144–151. doi: 10.1128/jb.121.1.144-151.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. 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]
  7. Harms E., Kern H., Schneider J. A. Human lysosomes can be purified from diploid skin fibroblasts by free-flow electrophoresis. Proc Natl Acad Sci U S A. 1980 Oct;77(10):6139–6143. doi: 10.1073/pnas.77.10.6139. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  9. Orloff S., Mukherjee A. B., Butler J. D., Foley B., Schulman J. D. Cystinotic and normal fibroblasts: differential susceptibility to cysteine toxicity in vitro. In Vitro. 1980 Aug;16(8):655–660. doi: 10.1007/BF02619194. [DOI] [PubMed] [Google Scholar]
  10. Oshima R. G., Willis R. C., Furlong C. E., Schneider J. A. Binding assays for amino acids. The utilization of a cystine binding protein from Escherichia coli for the determination of acid-soluble cystine in small physiological samples. J Biol Chem. 1974 Oct 10;249(19):6033–6039. [PubMed] [Google Scholar]
  11. Schneider J. A., Verroust F. M., Kroll W. A., Garvin A. J., Horger E. O., 3rd, Wong V. G., Spear G. S., Jacobson C., Pellett O. L., Becker F. L. Prenatal diagnosis of cystinosis. N Engl J Med. 1974 Apr 18;290(16):878–882. doi: 10.1056/NEJM197404182901604. [DOI] [PubMed] [Google Scholar]
  12. Thoene J. G., Oshima R. G., Crawhall J. C., Olson D. L., Schneider J. A. Cystinosis. Intracellular cystine depletion by aminothiols in vitro and in vivo. J Clin Invest. 1976 Jul;58(1):180–189. doi: 10.1172/JCI108448. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES