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. 1987 Mar 1;165(3):628–640. doi: 10.1084/jem.165.3.628

Induction of cystine transport activity in mouse peritoneal macrophages

PMCID: PMC2188288  PMID: 2880923

Abstract

Uptake of cystine was investigated in mouse peritoneal macrophages. The rates of the uptake of cystine in resident macrophages or macrophages elicited by some irritants were very low, but a drastic increase was observed when the cells were cultured in vitro. This increase was time- dependent and required protein synthesis. In macrophages elicited by thioglycollate broth, the rate of the uptake of cystine increased by about 40-fold after 16 h in culture. Contrary to the uptake of cystine, the rates of uptake of some neutral amino acids did not change markedly during culture. We characterized the induced activity of the cystine uptake in macrophages elicited by thioglycollate broth. Cystine was taken up in an Na+-independent and pH-sensitive manner, and the uptake was potently inhibited by extracellular glutamate and the analogous anionic amino acids, but not by aspartate. The activity of the glutamate uptake was also induced during the culture in a way similar to that of cystine uptake, and the uptake of glutamate was potently inhibited by cystine. From these results we concluded that the uptake of cystine and glutamate in macrophages was mostly mediated by a single transport system similar to the ones previously reported in human fibroblasts and some other cells. As a consequence of the induction of the activity of the cystine uptake, glutathione levels in macrophages doubled during culture, and a thiol compound, presumably cysteine, was released into the culture medium and accumulated there. When the macrophages were cultured hypoxically, the induction of the cystine uptake activity was markedly depressed, suggesting an involvement of oxygen in the induction.

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Selected References

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

  1. Bannai S. Exchange of cystine and glutamate across plasma membrane of human fibroblasts. J Biol Chem. 1986 Feb 15;261(5):2256–2263. [PubMed] [Google Scholar]
  2. Bannai S., Ishii T. Formation of sulfhydryl groups in the culture medium by human diploid fibroblasts. J Cell Physiol. 1980 Aug;104(2):215–223. doi: 10.1002/jcp.1041040211. [DOI] [PubMed] [Google Scholar]
  3. Bannai S., Ishii T. Transport of cystine and cysteine and cell growth in cultured human diploid fibroblasts: effect of glutamate and homocysteate. J Cell Physiol. 1982 Aug;112(2):265–272. doi: 10.1002/jcp.1041120216. [DOI] [PubMed] [Google Scholar]
  4. Bannai S., Kitamura E. Role of proton dissociation in the transport of cystine and glutamate in human diploid fibroblasts in culture. J Biol Chem. 1981 Jun 10;256(11):5770–5772. [PubMed] [Google Scholar]
  5. Bannai S., Kitamura E. Transport interaction of L-cystine and L-glutamate in human diploid fibroblasts in culture. J Biol Chem. 1980 Mar 25;255(6):2372–2376. [PubMed] [Google Scholar]
  6. Bannai S., Tateishi N. Role of membrane transport in metabolism and function of glutathione in mammals. J Membr Biol. 1986;89(1):1–8. doi: 10.1007/BF01870891. [DOI] [PubMed] [Google Scholar]
  7. Christensen H. N. Exploiting amino acid structure to learn about membrane transport. Adv Enzymol Relat Areas Mol Biol. 1979;49:41–101. doi: 10.1002/9780470122945.ch2. [DOI] [PubMed] [Google Scholar]
  8. Fedorcsák I., Harms-Ringdahl M., Ehrenberg L. Prevention of sulfhydryl autoxidation by a polypeptide from red kidney beans, described to be a stimulator of RNA synthesis. Exp Cell Res. 1977 Sep;108(2):331–339. doi: 10.1016/s0014-4827(77)80040-2. [DOI] [PubMed] [Google Scholar]
  9. Hishinuma I., Ishii T., Watanabe H., Bannai S. Mouse lymphoma L1210 cells acquire a new cystine transport activity upon adaptation in vitro. In Vitro Cell Dev Biol. 1986 Mar;22(3 Pt 1):127–134. doi: 10.1007/BF02623499. [DOI] [PubMed] [Google Scholar]
  10. Ishii T., Bannai S., Sugita Y. Mechanism of growth stimulation of L1210 cells by 2-mercaptoethanol in vitro. Role of the mixed disulfide of 2-mercaptoethanol and cysteine. J Biol Chem. 1981 Dec 10;256(23):12387–12392. [PubMed] [Google Scholar]
  11. Ishii T., Hishinuma I., Bannai S., Sugita Y. Mechanism of growth promotion of mouse lymphoma L1210 cells in vitro by feeder layer or 2-mercaptoethanol. J Cell Physiol. 1981 May;107(2):283–293. doi: 10.1002/jcp.1041070215. [DOI] [PubMed] [Google Scholar]
  12. Makowske M., Christensen H. N. Contrasts in transport systems for anionic amino acids in hepatocytes and a hepatoma cell line HTC. J Biol Chem. 1982 May 25;257(10):5663–5670. [PubMed] [Google Scholar]
  13. Nathan C. F., Terry W. D. Differential stimulation of murine lymphoma growth in vitro by normal and BCG-activated macrophages. J Exp Med. 1975 Oct 1;142(4):887–902. doi: 10.1084/jem.142.4.887. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Novogrodsky A., Nehring R. E., Jr, Meister A. Inhibition of amino acid transport into lymphoid cells by the glutamine analog L-2-amino-4-oxo-5-chloropentanoate. Proc Natl Acad Sci U S A. 1979 Oct;76(10):4932–4935. doi: 10.1073/pnas.76.10.4932. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Ohmori H., Yamamoto I. Mechanism of augmentation of the antibody response in vitro by 2-mercaptoethanol in murine lymphocytes. I. 2-Mercaptoethanol-induced stimulation of the uptake of cystine, an essential amino acid. J Exp Med. 1982 May 1;155(5):1277–1290. doi: 10.1084/jem.155.5.1277. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Rouzer C. A., Scott W. A., Griffith O. W., Hamill A. L., Cohn Z. A. Depletion of glutathione selectively inhibits synthesis of leukotriene C by macrophages. Proc Natl Acad Sci U S A. 1981 Apr;78(4):2532–2536. doi: 10.1073/pnas.78.4.2532. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Rouzer C. A., Scott W. A., Griffith O. W., Hamill A. L., Cohn Z. A. Glutathione metabolism in resting and phagocytizing peritoneal macrophages. J Biol Chem. 1982 Feb 25;257(4):2002–2008. [PubMed] [Google Scholar]
  18. Rouzer C. A., Scott W. A., Hamill A. L., Liu F. T., Katz D. H., Cohn Z. A. Secretion of leukotriene C and other arachidonic acid metabolites by macrophages challenged with immunoglobulin E immune complexes. J Exp Med. 1982 Oct 1;156(4):1077–1086. doi: 10.1084/jem.156.4.1077. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Steinherz R., Tietze F., Raiford D., Gahl W. A., Schulman J. D. Patterns of amino acid efflux from isolated normal and cystinotic human leucocyte lysosomes. J Biol Chem. 1982 Jun 10;257(11):6041–6049. [PubMed] [Google Scholar]
  20. Takada A., Bannai S. Transport of cystine in isolated rat hepatocytes in primary culture. J Biol Chem. 1984 Feb 25;259(4):2441–2445. [PubMed] [Google Scholar]
  21. Tanapet P., Gaetjens E., Broome J. D. Growth promoting and inhibitory activities of 3T3 and other cell lines for thioldependent lymphoma cells in vitro. Proc Soc Exp Biol Med. 1978 Mar;157(3):517–521. doi: 10.3181/00379727-157-40088. [DOI] [PubMed] [Google Scholar]
  22. Tietze F. Enzymic method for quantitative determination of nanogram amounts of total and oxidized glutathione: applications to mammalian blood and other tissues. Anal Biochem. 1969 Mar;27(3):502–522. doi: 10.1016/0003-2697(69)90064-5. [DOI] [PubMed] [Google Scholar]

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