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. 1986 Jan;83(1):150–154. doi: 10.1073/pnas.83.1.150

Functional channel formation associated with cytotoxic T-cell granules.

J D Young, C F Nathan, E R Podack, M A Palladino, Z A Cohn
PMCID: PMC322809  PMID: 2417234

Abstract

Lymphocyte granules from cytotoxic T-lymphocyte lines A2, A11, and R8 were enriched by subcellular fractionation using a Percoll gradient. Granule-enriched fractions showed potent hemolytic activity in the presence of Ca2+. Isolated granules induced rapid Ca2+-dependent membrane depolarization of J774 macrophage-like cells. When tested in planar bilayers, granules induced the formation of Ca2+-dependent functional ion channels of large conductance steps of 1-6 nS in 0.1 M NaCl. Granule-induced channels were resistant to closing by an increase in transmembrane potential, with few channels shifting to the closed state only at voltages of greater than 70 mV, following a Poisson process. These channels showed poor ion selectivity and were permeable to all monovalent and divalent ions (K+, Na+, Li+, Cl-, Ca2+, Mg2+, Zn2+, Ba2+). Ultrastructural examination of soluble granule proteins incubated for 48 hr at 37 degrees C in the presence of Ca2+ revealed ring-like structures of 150-200 A. Structural and functional channel formation may be involved in cytolysis induced by cytotoxic T lymphocytes.

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Selected References

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

  1. Borregaard N., Heiple J. M., Simons E. R., Clark R. A. Subcellular localization of the b-cytochrome component of the human neutrophil microbicidal oxidase: translocation during activation. J Cell Biol. 1983 Jul;97(1):52–61. doi: 10.1083/jcb.97.1.52. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Dennert G., Podack E. R. Cytolysis by H-2-specific T killer cells. Assembly of tubular complexes on target membranes. J Exp Med. 1983 May 1;157(5):1483–1495. doi: 10.1084/jem.157.5.1483. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Dourmashkin R. R., Deteix P., Simone C. B., Henkart P. Electron microscopic demonstration of lesions in target cell membranes associated with antibody-dependent cellular cytotoxicity. Clin Exp Immunol. 1980 Dec;42(3):554–560. [PMC free article] [PubMed] [Google Scholar]
  4. Henkart P. A., Millard P. J., Reynolds C. W., Henkart M. P. Cytolytic activity of purified cytoplasmic granules from cytotoxic rat large granular lymphocyte tumors. J Exp Med. 1984 Jul 1;160(1):75–93. doi: 10.1084/jem.160.1.75. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Latorre R., Alvarez O. Voltage-dependent channels in planar lipid bilayer membranes. Physiol Rev. 1981 Jan;61(1):77–150. doi: 10.1152/physrev.1981.61.1.77. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. Palladino M. A., Obata Y., Stockert E., Oettgen H. F. Characterization of interleukin 2-dependent cytotoxic T-cell clones: specificity, cell surface phenotype, and susceptibility to blocking by Lyt antisera. Cancer Res. 1983 Feb;43(2):572–576. [PubMed] [Google Scholar]
  8. Podack E. R., Dennert G. Assembly of two types of tubules with putative cytolytic function by cloned natural killer cells. 1983 Mar 31-Apr 6Nature. 302(5907):442–445. doi: 10.1038/302442a0. [DOI] [PubMed] [Google Scholar]
  9. Podack E. R., Konigsberg P. J. Cytolytic T cell granules. Isolation, structural, biochemical, and functional characterization. J Exp Med. 1984 Sep 1;160(3):695–710. doi: 10.1084/jem.160.3.695. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Podack E. R., Tschopp J. Circular polymerization of the ninth component of complement. Ring closure of the tubular complex confers resistance to detergent dissociation and to proteolytic degradation. J Biol Chem. 1982 Dec 25;257(24):15204–15212. [PubMed] [Google Scholar]
  11. Schein S. J., Kagan B. L., Finkelstein A. Colicin K acts by forming voltage-dependent channels in phospholipid bilayer membranes. Nature. 1978 Nov 9;276(5684):159–163. doi: 10.1038/276159a0. [DOI] [PubMed] [Google Scholar]
  12. Tschopp J., Müller-Eberhard H. J., Podack E. R. Formation of transmembrane tubules by spontaneous polymerization of the hydrophilic complement protein C9. Nature. 1982 Aug 5;298(5874):534–538. doi: 10.1038/298534a0. [DOI] [PubMed] [Google Scholar]
  13. Young G. P., Young J. D., Deshpande A. K., Goldstein M., Koide S. S., Cohn Z. A. A Ca2+-activated channel from Xenopus laevis oocyte membranes reconstituted into planar bilayers. Proc Natl Acad Sci U S A. 1984 Aug;81(16):5155–5159. doi: 10.1073/pnas.81.16.5155. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Young J. D., Blake M., Mauro A., Cohn Z. A. Properties of the major outer membrane protein from Neisseria gonorrhoeae incorporated into model lipid membranes. Proc Natl Acad Sci U S A. 1983 Jun;80(12):3831–3835. doi: 10.1073/pnas.80.12.3831. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Young J. D., Unkeless J. C., Kaback H. R., Cohn Z. A. Macrophage membrane potential changes associated with gamma 2b/gamma 1 Fc receptor-ligand binding. Proc Natl Acad Sci U S A. 1983 Mar;80(5):1357–1361. doi: 10.1073/pnas.80.5.1357. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Young J. D., Unkeless J. C., Young T. M., Mauro A., Cohn Z. A. Role for mouse macrophage IgG Fc receptor as ligand-dependent ion channel. Nature. 1983 Nov 10;306(5939):186–189. doi: 10.1038/306186a0. [DOI] [PubMed] [Google Scholar]
  17. Young J. D., Young T. M., Lu L. P., Unkeless J. C., Cohn Z. A. Characterization of a membrane pore-forming protein from Entamoeba histolytica. J Exp Med. 1982 Dec 1;156(6):1677–1690. doi: 10.1084/jem.156.6.1677. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Young T. M., Young J. D. Protein-mediated intermembrane contact facilitates fusion of lipid vesicles with planar bilayers. Biochim Biophys Acta. 1984 Sep 5;775(3):441–445. doi: 10.1016/0005-2736(84)90202-5. [DOI] [PubMed] [Google Scholar]

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