Skip to main content
NCBI 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
. 1989 Sep;86(18):7128–7132. doi: 10.1073/pnas.86.18.7128

Two proteins targeted to the same lytic granule compartment undergo very different posttranslational processing.

J K Burkhardt 1, S Hester 1, Y Argon 1
PMCID: PMC298008  PMID: 2674947

Abstract

The granules of natural killer (NK) cells contain cytolysin and serine proteases, proteins that are expressed specifically in cytolytic cells and are released in response to target binding. We have used immunofluorescence and immunoelectron microscopy to localize these proteins with respect to the various morphological compartments of granules in a rat NK cell line. Both cytolysin and the proteases are limited to the core regions of the dense core granules. While these proteins are targeted to the same compartment, they differ markedly in their posttranslational processing. Cytolysin bears N-linked oligosaccharides that are converted to the complex type, while the major trypsin-like protease, granzyme A, bears only high-mannose-type oligosaccharides. The glycans of granzyme A, but not those of cytolysin, are modified with phosphomannose moieties. These results suggest that one possible mechanism for packaging proteins into NK granules is the mannose 6-phosphate-dependent lysosomal targeting system. However, the absence of the mannose 6-phosphate modification from cytolysin suggests the existence of yet another targeting system.

Full text

PDF
7128

Images in this article

7129Image
on p.7129
7130Image
on p.7130
7130Image
on p.7130
7131Image
on p.7131
7131Image
on p.7131

Selected References

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

  1. Argon Y., Milstein C. Intracellular processing of membrane and secreted immunoglobulin delta-chains. J Immunol. 1984 Sep;133(3):1627–1634. [PubMed] [Google Scholar]
  2. Bole D. G., Hendershot L. M., Kearney J. F. Posttranslational association of immunoglobulin heavy chain binding protein with nascent heavy chains in nonsecreting and secreting hybridomas. J Cell Biol. 1986 May;102(5):1558–1566. doi: 10.1083/jcb.102.5.1558. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brown W. J., Farquhar M. G. The mannose-6-phosphate receptor for lysosomal enzymes is concentrated in cis Golgi cisternae. Cell. 1984 Feb;36(2):295–307. doi: 10.1016/0092-8674(84)90223-x. [DOI] [PubMed] [Google Scholar]
  4. Burgess T. L., Kelly R. B. Constitutive and regulated secretion of proteins. Annu Rev Cell Biol. 1987;3:243–293. doi: 10.1146/annurev.cb.03.110187.001331. [DOI] [PubMed] [Google Scholar]
  5. Burkhardt J. K., Hester S., Argon Y. The glycoprotein of VSV accumulates in a distal Golgi compartment in the presence of CCCP. J Cell Sci. 1989 Apr;92(Pt 4):643–654. doi: 10.1242/jcs.92.4.643. [DOI] [PubMed] [Google Scholar]
  6. Distler J., Hieber V., Sahagian G., Schmickel R., Jourdian G. W. Identification of mannose 6-phosphate in glycoproteins that inhibit the assimilation of beta-galactosidase by fibroblasts. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4235–4239. doi: 10.1073/pnas.76.9.4235. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dvorak A. M., Galli S. J., Marcum J. A., Nabel G., der Simonian H., Goldin J., Monahan R. A., Pyne K., Cantor H., Rosenberg R. D. Cloned mouse cells with natural killer function and cloned suppressor T cells express ultrastructural and biochemical features not shared by cloned inducer T cells. J Exp Med. 1983 Mar 1;157(3):843–861. doi: 10.1084/jem.157.3.843. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Frey T., Petty H. R., McConnell H. M. Electron microscopic study of natural killer cell-tumor cell conjugates. Proc Natl Acad Sci U S A. 1982 Sep;79(17):5317–5321. doi: 10.1073/pnas.79.17.5317. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gershenfeld H. K., Weissman I. L. Cloning of a cDNA for a T cell-specific serine protease from a cytotoxic T lymphocyte. Science. 1986 May 16;232(4752):854–858. doi: 10.1126/science.2422755. [DOI] [PubMed] [Google Scholar]
  10. Griffiths G., Simons K. The trans Golgi network: sorting at the exit site of the Golgi complex. Science. 1986 Oct 24;234(4775):438–443. doi: 10.1126/science.2945253. [DOI] [PubMed] [Google Scholar]
  11. Groscurth P., Qiao B. Y., Podack E. R., Hengartner H. Cellular localization of perforin 1 in murine cloned cytotoxic T lymphocytes. J Immunol. 1987 May 1;138(9):2749–2752. [PubMed] [Google Scholar]
  12. Henkart P. A., Berrebi G. A., Takayama H., Munger W. E., Sitkovsky M. V. Biochemical and functional properties of serine esterases in acidic cytoplasmic granules of cytotoxic T lymphocytes. J Immunol. 1987 Oct 1;139(7):2398–2405. [PubMed] [Google Scholar]
  13. Henkart P. A. Mechanism of lymphocyte-mediated cytotoxicity. Annu Rev Immunol. 1985;3:31–58. doi: 10.1146/annurev.iy.03.040185.000335. [DOI] [PubMed] [Google Scholar]
  14. Jenne D., Rey C., Haefliger J. A., Qiao B. Y., Groscurth P., Tschopp J. Identification and sequencing of cDNA clones encoding the granule-associated serine proteases granzymes D, E, and F of cytolytic T lymphocytes. Proc Natl Acad Sci U S A. 1988 Jul;85(13):4814–4818. doi: 10.1073/pnas.85.13.4814. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kornfeld R., Kornfeld S. Assembly of asparagine-linked oligosaccharides. Annu Rev Biochem. 1985;54:631–664. doi: 10.1146/annurev.bi.54.070185.003215. [DOI] [PubMed] [Google Scholar]
  16. Kozutsumi Y., Segal M., Normington K., Gething M. J., Sambrook J. The presence of malfolded proteins in the endoplasmic reticulum signals the induction of glucose-regulated proteins. Nature. 1988 Mar 31;332(6163):462–464. doi: 10.1038/332462a0. [DOI] [PubMed] [Google Scholar]
  17. Kupfer A., Dennert G., Singer S. J. Polarization of the Golgi apparatus and the microtubule-organizing center within cloned natural killer cells bound to their targets. Proc Natl Acad Sci U S A. 1983 Dec;80(23):7224–7228. doi: 10.1073/pnas.80.23.7224. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Lobe C. G., Finlay B. B., Paranchych W., Paetkau V. H., Bleackley R. C. Novel serine proteases encoded by two cytotoxic T lymphocyte-specific genes. Science. 1986 May 16;232(4752):858–861. doi: 10.1126/science.3518058. [DOI] [PubMed] [Google Scholar]
  19. MacDermott R. P., Schmidt R. E., Caulfield J. P., Hein A., Bartley G. T., Ritz J., Schlossman S. F., Austen K. F., Stevens R. L. Proteoglycans in cell-mediated cytotoxicity. Identification, localization, and exocytosis of a chondroitin sulfate proteoglycan from human cloned natural killer cells during target cell lysis. J Exp Med. 1985 Dec 1;162(6):1771–1787. doi: 10.1084/jem.162.6.1771. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Masson D., Tschopp J. A family of serine esterases in lytic granules of cytolytic T lymphocytes. Cell. 1987 Jun 5;49(5):679–685. doi: 10.1016/0092-8674(87)90544-7. [DOI] [PubMed] [Google Scholar]
  21. Millard P. J., Henkart M. P., Reynolds C. W., Henkart P. A. Purification and properties of cytoplasmic granules from cytotoxic rat LGL tumors. J Immunol. 1984 Jun;132(6):3197–3204. [PubMed] [Google Scholar]
  22. Neighbour P. A., Huberman H. S., Kress Y. Human large granular lymphocytes and natural killing ultrastructural studies of strontium-induced degranulation. Eur J Immunol. 1982 Jul;12(7):588–595. doi: 10.1002/eji.1830120711. [DOI] [PubMed] [Google Scholar]
  23. Orye E., Plum J., De Smedt M. Beta-glucuronidase activity in human T and B lymphocytes and the Tmu and T gamma subpopulations. Histochemistry. 1984;81(3):287–290. doi: 10.1007/BF00495641. [DOI] [PubMed] [Google Scholar]
  24. Pasternack M. S., Eisen H. N. A novel serine esterase expressed by cytotoxic T lymphocytes. 1985 Apr 25-May 1Nature. 314(6013):743–745. doi: 10.1038/314743a0. [DOI] [PubMed] [Google Scholar]
  25. Podack E. R., Young J. D., Cohn Z. A. Isolation and biochemical and functional characterization of perforin 1 from cytolytic T-cell granules. Proc Natl Acad Sci U S A. 1985 Dec;82(24):8629–8633. doi: 10.1073/pnas.82.24.8629. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Reynolds C. W., Bere E. W., Jr, Ward J. M. Natural killer activity in the rat. III. Characterization of transplantable large granular lymphocyte (LGL) leukemias in the F344 rat. J Immunol. 1984 Jan;132(1):534–540. [PubMed] [Google Scholar]
  27. Reynolds C. W., Reichardt D., Henkart M., Millard P., Henkart P. Inhibition of NK and ADCC activity by antibodies against purified cytoplasmic granules from rat LGL tumors. J Leukoc Biol. 1987 Dec;42(6):642–652. doi: 10.1002/jlb.42.6.642. [DOI] [PubMed] [Google Scholar]
  28. Roth J., Berger E. G. Immunocytochemical localization of galactosyltransferase in HeLa cells: codistribution with thiamine pyrophosphatase in trans-Golgi cisternae. J Cell Biol. 1982 Apr;93(1):223–229. doi: 10.1083/jcb.93.1.223. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Takatsu K., Sano Y., Hashimoto N., Tomita S., Hamaoka T. Acceptor site(s) for T cell-replacing factor (TRF) on B lymphocytes. I. TRF-substituting activity of anti-TRF acceptor site(s) antibody in the triggering of B cells. J Immunol. 1982 Jun;128(6):2575–2580. [PubMed] [Google Scholar]
  30. Takayama H., Trenn G., Humphrey W., Jr, Bluestone J. A., Henkart P. A., Sitkovsky M. V. Antigen receptor-triggered secretion of a trypsin-type esterase from cytotoxic T lymphocytes. J Immunol. 1987 Jan 15;138(2):566–569. [PubMed] [Google Scholar]
  31. Zagury D. Direct analysis of individual killer T cells: susceptibility of target cells to lysis and secretion of hydrolytic enzymes by CTL. Adv Exp Med Biol. 1982;146:149–169. doi: 10.1007/978-1-4684-8959-0_10. [DOI] [PubMed] [Google Scholar]
  32. Zucker-Franklin D., Grusky G., Yang J. S. Arylsulfatase in natural killer cells: its possible role in cytotoxicity. Proc Natl Acad Sci U S A. 1983 Nov;80(22):6977–6981. doi: 10.1073/pnas.80.22.6977. [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