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. 1970 Nov 30;132(6):1090–1104. doi: 10.1084/jem.132.6.1090

THE METABOLISM OF AMINO ACIDS, PEPTIDES, AND DISULFIDES IN LYSOSOMES OF FIBROBLASTS CULTURED FROM NORMAL INDIVIDUALS AND THOSE WITH CYSTINOSIS

Joseph D Schulman 1, Kathryn H Bradley 1
PMCID: PMC2180497  PMID: 5516432

Abstract

The metabolism of amino acids, peptides, and disulfides has been investigates in cultured skin fibroblasts from normal individuals and patients with cystinosis. Human fibroblast lysosomes closely resemble the lysosomes of mouse peritoneal macrophages in having an apparent permeability barrier to amino acids and peptides with molecular weights of greater than 220–230. Cystinotic and normal cells behave similarly in this regard. Normal cells do not undergo lysosomal swelling when exposed to cysteine-penicillamine disulfides, while cystinotic cells are prominently vacuolized under these conditions. Normal lysosomes may have a specific mechanism for the disposal of cystine, and deficient activity of this mechanism in cystinotic lysosomes could result in cystine storage therein. The demonstration that human fibroblasts can be used conveniently to study lysosomal metabolism of small substrates may facilitate investigations of these aspects of lysosomal function in a variety of genetic diseases of man.

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Selected References

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

  1. Brady R. O., Gal A. E., Bradley R. M., Martensson E., Warshaw A. L., Laster L. Enzymatic defect in Fabry's disease. Ceramidetrihexosidase deficiency. N Engl J Med. 1967 May 25;276(21):1163–1167. doi: 10.1056/NEJM196705252762101. [DOI] [PubMed] [Google Scholar]
  2. Brady R. O., Kanfer J. N., Bradley R. M., Shapiro D. Demonstration of a deficiency of glucocerebroside-cleaving enzyme in Gaucher's disease. J Clin Invest. 1966 Jul;45(7):1112–1115. doi: 10.1172/JCI105417. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brady R. O., Kanfer J. N., Mock M. B., Fredrickson D. S. The metabolism of sphingomyelin. II. Evidence of an enzymatic deficiency in Niemann-Pick diseae. Proc Natl Acad Sci U S A. 1966 Feb;55(2):366–369. doi: 10.1073/pnas.55.2.366. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. COGAN D. G., KUWABARA T., KINOSHITA J., SHEEHAN L., MEROLA L. Cystinosis in an adult. J Am Med Assoc. 1957 May 25;164(4):394–396. doi: 10.1001/jama.1957.02980040034009. [DOI] [PubMed] [Google Scholar]
  5. CRAWHALL J. C., SCOWEN E. F., WATTS R. W. FURTHER OBSERVATIONS ON USE OF D-PENICILLAMINE IN CYSTINURIA. Br Med J. 1964 May 30;1(5395):1411–1413. doi: 10.1136/bmj.1.5395.1411. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chambers R. W., Bowling M. C., Grimley P. M. Glutaraldehyde fixation in routine histopathology. Arch Pathol. 1968 Jan;85(1):18–30. [PubMed] [Google Scholar]
  7. Crawhall J. C., Thompson C. J., Bradley K. H. Separation of cystine, penicillamine disulfide and cysteine-penicillamine mixed disulfide by automatic amino acid analysis. Anal Biochem. 1966 Mar;14(3):405–413. doi: 10.1016/0003-2697(66)90282-x. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. Goldman H., Scriver C. R., Aaron K., Pinsky L. Use of dithiothreitol to correct cystine storage in cultured cystinotic fibroblasts. Lancet. 1970 Apr 18;1(7651):811–812. doi: 10.1016/s0140-6736(70)92411-6. [DOI] [PubMed] [Google Scholar]
  10. Hegner D. Kationenabhängige Membran-ATPasen in lysosomalen granula aus polymorphkernigen Rinderleukocyten. Naunyn Schmiedebergs Arch Exp Pathol Pharmakol. 1969;263(3):406–416. [PubMed] [Google Scholar]
  11. Hummeler K., Zajac B. A., Genel M., Holtzapple P. G., Segal S. Human cystinosis: intracellular deposition of cystine. Science. 1970 May 15;168(3933):859–860. doi: 10.1126/science.168.3933.859. [DOI] [PubMed] [Google Scholar]
  12. Lake B. D., Patrick A. D. Wolman's disease: deficiency of E600-resistant acid esterase activity with storage of lipids in lysosomes. J Pediatr. 1970 Feb;76(2):262–266. doi: 10.1016/s0022-3476(70)80172-x. [DOI] [PubMed] [Google Scholar]
  13. Lieb W. R., Stein W. D. Biological membranes behave as non-porous polymeric sheets with respect to the diffusion of non-electrolytes. Nature. 1969 Oct 18;224(5216):240–243. doi: 10.1038/224240a0. [DOI] [PubMed] [Google Scholar]
  14. Lietman P. S., Frazier P. D., Wong V. G., Shotton D., Seegmiller J. E. Adult cystinosis--a benign disorder. Am J Med. 1966 Apr;40(4):511–517. doi: 10.1016/0002-9343(66)90115-x. [DOI] [PubMed] [Google Scholar]
  15. MEHL E., JATZKEWITZ H. EVIDENCE FOR THE GENETIC BLOCK IN METACHROMATIC LEUCODYSTROPHY (ML). Biochem Biophys Res Commun. 1965 May 3;19:407–411. doi: 10.1016/0006-291x(65)90137-3. [DOI] [PubMed] [Google Scholar]
  16. Morecki R., Paunier L., Hamilton J. R. Intestinal mucosa in cystinosis. A fine structure study. Arch Pathol. 1968 Sep;86(3):297–307. [PubMed] [Google Scholar]
  17. Nadler H. L., Egan T. J. Deficiency of lysosomal acid phosphatase. A new familial metabolic disorder. N Engl J Med. 1970 Feb 5;282(6):302–307. doi: 10.1056/NEJM197002052820604. [DOI] [PubMed] [Google Scholar]
  18. Okada S., O'Brien J. S. Generalized gangliosidosis: beta-galactosidase deficiency. Science. 1968 May 31;160(3831):1002–1004. doi: 10.1126/science.160.3831.1002. [DOI] [PubMed] [Google Scholar]
  19. Okada S., O'Brien J. S. Tay-Sachs disease: generalized absence of a beta-D-N-acetylhexosaminidase component. Science. 1969 Aug 15;165(3894):698–700. doi: 10.1126/science.165.3894.698. [DOI] [PubMed] [Google Scholar]
  20. 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]
  21. Schneider J. A., Bradley K., Seegmiller J. E. Increased cystine in leukocytes from individuals homozygous and heterozygous for cystinosis. Science. 1967 Sep 15;157(3794):1321–1322. doi: 10.1126/science.157.3794.1321. [DOI] [PubMed] [Google Scholar]
  22. 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]
  23. 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]
  24. Schulman J. D., Fujimoto W. Y., Bradley K. H., Seegmiller J. E. Identification of heterozygous genotype for cystinosis in utero by a new pulse-labeling technique: preliminary report. J Pediatr. 1970 Sep;77(3):468–470. doi: 10.1016/s0022-3476(70)80020-8. [DOI] [PubMed] [Google Scholar]
  25. Schulman J. D., Wong V., Olson W. H., Seegmiller J. E. Lysosomal site of crystalline deposits in cystinosis as shown by ferritin uptake. Arch Pathol. 1970 Sep;90(3):259–264. [PubMed] [Google Scholar]
  26. Seegmiller J. E., Friedmann T., Harrison H. E., Wong V., Schneider J. A. Cystinosis. Combined clinical staff conference at the National Institutes of Health. Ann Intern Med. 1968 Apr;68(4):883–905. doi: 10.7326/0003-4819-68-4-883. [DOI] [PubMed] [Google Scholar]
  27. States B., Segal S. Distribution of glutathione-cystine transhydrogenase activity in subcellular fractions of rat intestinal mucosa. Biochem J. 1969 Jun;113(2):443–444. doi: 10.1042/bj1130443. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Van Hoof F., Hers H. G. Mucopolysaccharidosis by absence of alpha-fucosidase. Lancet. 1968 Jun 1;1(7553):1198–1198. doi: 10.1016/s0140-6736(68)91895-3. [DOI] [PubMed] [Google Scholar]
  29. Wong V. G., Kuwabara T., Brubaker R., Olson W., Schulman J., Seegmiller J. E. Intralysosomal cystine crystals in cystinosis. Invest Ophthalmol. 1970 Jan;9(1):83–88. [PubMed] [Google Scholar]
  30. Woodin A. M., Wieneke A. A. The participation of calcium, adenosine triphosphate and adenosine triphosphatase in the extrusion of the granule proteins from the polymorphonuclear leucocyte. Biochem J. 1964 Mar;90(3):498–509. doi: 10.1042/bj0900498. [DOI] [PMC free article] [PubMed] [Google Scholar]

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