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. 1990 May 1;110(5):1525–1531. doi: 10.1083/jcb.110.5.1525

Dynamics and longevity of the glycolipid-anchored membrane protein, Thy- 1

PMCID: PMC2200196  PMID: 1970823

Abstract

Thy-1 and a number of other proteins are anchored to the outer hemi- leaflet of membranes by a glycolipid moiety containing ethanolamine phosphate, mannose, glucosamine, and phosphatidylinositol. They nevertheless have the striking property of being able to transduce signals across the plasma membrane. We here demonstrate, for the BW5147 murine T lymphoma, that (a) greater than 90% of Thy-1 is at the cell surface, (b) Thy-1 is about one order of magnitude less concentrated in coated pits than the transferrin receptor or H-2 antigens, (c) Thy-1 undergoes at most very limited endocytosis or diacytosis, and (d) Thy-1 has an unusually slow turnover rate. Several similar observations have also been made for a second glycolipid-anchored protein, the T cell activating protein. Thus, the absence of cytoplasmic and trans-membrane domains may result in lipid-anchored proteins being confined to the cell surface and being free from constraints which affect the turnover of transmembrane proteins.

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Selected References

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  1. Bamezai A., Goldmacher V., Reiser H., Rock K. L. Internalization of phosphatidylinositol-anchored lymphocyte proteins. I. Documentation and potential significance for T cell stimulation. J Immunol. 1989 Nov 15;143(10):3107–3116. [PubMed] [Google Scholar]
  2. Bamezai A., Reiser H., Rock K. L. T cell receptor/CD3 negative variants are unresponsive to stimulation through the Ly-6 encoded molecule, TAP. J Immunol. 1988 Sep 1;141(5):1423–1428. [PubMed] [Google Scholar]
  3. Baron M. D., Luzio J. P. The synthesis and turnover of 5'-nucleotidase in primary cultured hepatocytes. Biochim Biophys Acta. 1987 Jan 19;927(1):81–85. doi: 10.1016/0167-4889(87)90068-1. [DOI] [PubMed] [Google Scholar]
  4. Bretscher M. S., Thomson J. N., Pearse B. M. Coated pits act as molecular filters. Proc Natl Acad Sci U S A. 1980 Jul;77(7):4156–4159. doi: 10.1073/pnas.77.7.4156. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Brown W. J., Farquhar M. G. Immunoperoxidase methods for the localization of antigens in cultured cells and tissue sections by electron microscopy. Methods Cell Biol. 1989;31:553–569. doi: 10.1016/s0091-679x(08)61626-x. [DOI] [PubMed] [Google Scholar]
  6. Bülow R., Overath P., Davoust J. Rapid lateral diffusion of the variant surface glycoprotein in the coat of Trypanosoma brucei. Biochemistry. 1988 Apr 5;27(7):2384–2388. doi: 10.1021/bi00407a020. [DOI] [PubMed] [Google Scholar]
  7. Campbell D. G., Gagnon J., Reid K. B., Williams A. F. Rat brain Thy-1 glycoprotein. The amino acid sequence, disulphide bonds and an unusual hydrophobic region. Biochem J. 1981 Apr 1;195(1):15–30. doi: 10.1042/bj1950015. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Chan B. L., Lisanti M. P., Rodriguez-Boulan E., Saltiel A. R. Insulin-stimulated release of lipoprotein lipase by metabolism of its phosphatidylinositol anchor. Science. 1988 Sep 23;241(4873):1670–1672. doi: 10.1126/science.241.4873.1670. [DOI] [PubMed] [Google Scholar]
  9. Conzelmann A., Riezman H., Desponds C., Bron C. A major 125-kd membrane glycoprotein of Saccharomyces cerevisiae is attached to the lipid bilayer through an inositol-containing phospholipid. EMBO J. 1988 Jul;7(7):2233–2240. doi: 10.1002/j.1460-2075.1988.tb03063.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Conzelmann A., Spiazzi A., Bron C., Hyman R. No glycolipid anchors are added to Thy-1 glycoprotein in Thy-1-negative mutant thymoma cells of four different complementation classes. Mol Cell Biol. 1988 Feb;8(2):674–678. doi: 10.1128/mcb.8.2.674. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Davignon D., Martz E., Reynolds T., Kürzinger K., Springer T. A. Lymphocyte function-associated antigen 1 (LFA-1): a surface antigen distinct from Lyt-2,3 that participates in T lymphocyte-mediated killing. Proc Natl Acad Sci U S A. 1981 Jul;78(7):4535–4539. doi: 10.1073/pnas.78.7.4535. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Davis C. G., van Driel I. R., Russell D. W., Brown M. S., Goldstein J. L. The low density lipoprotein receptor. Identification of amino acids in cytoplasmic domain required for rapid endocytosis. J Biol Chem. 1987 Mar 25;262(9):4075–4082. [PubMed] [Google Scholar]
  13. Davis L. S., Patel S. S., Atkinson J. P., Lipsky P. E. Decay-accelerating factor functions as a signal transducing molecule for human T cells. J Immunol. 1988 Oct 1;141(7):2246–2252. [PubMed] [Google Scholar]
  14. Dragsten P., Henkart P., Blumenthal R., Weinstein J., Schlessinger J. Lateral diffusion of surface immunoglobulin, Thy-1 antigen, and a lipid probe in lymphocyte plasma membranes. Proc Natl Acad Sci U S A. 1979 Oct;76(10):5163–5167. doi: 10.1073/pnas.76.10.5163. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Fatemi S. H., Tartakoff A. M. The phenotype of five classes of T lymphoma mutants. Defective glycophospholipid anchoring, rapid degradation, and secretion of Thy-1 glycoprotein. J Biol Chem. 1988 Jan 25;263(3):1288–1294. [PubMed] [Google Scholar]
  16. Ferguson M. A., Williams A. F. Cell-surface anchoring of proteins via glycosyl-phosphatidylinositol structures. Annu Rev Biochem. 1988;57:285–320. doi: 10.1146/annurev.bi.57.070188.001441. [DOI] [PubMed] [Google Scholar]
  17. Hanover J. A., Willingham M. C., Pastan I. Kinetics of transit of transferrin and epidermal growth factor through clathrin-coated membranes. Cell. 1984 Dec;39(2 Pt 1):283–293. doi: 10.1016/0092-8674(84)90006-0. [DOI] [PubMed] [Google Scholar]
  18. Homans S. W., Ferguson M. A., Dwek R. A., Rademacher T. W., Anand R., Williams A. F. Complete structure of the glycosyl phosphatidylinositol membrane anchor of rat brain Thy-1 glycoprotein. Nature. 1988 May 19;333(6170):269–272. doi: 10.1038/333269a0. [DOI] [PubMed] [Google Scholar]
  19. Hopkins C. R., Trowbridge I. S. Internalization and processing of transferrin and the transferrin receptor in human carcinoma A431 cells. J Cell Biol. 1983 Aug;97(2):508–521. doi: 10.1083/jcb.97.2.508. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Huizinga T. W., van der Schoot C. E., Jost C., Klaassen R., Kleijer M., von dem Borne A. E., Roos D., Tetteroo P. A. The PI-linked receptor FcRIII is released on stimulation of neutrophils. Nature. 1988 Jun 16;333(6174):667–669. doi: 10.1038/333667a0. [DOI] [PubMed] [Google Scholar]
  21. Hämmerling G. J. T lymphocyte tissue culture lines produced by cell hybridization. Eur J Immunol. 1977 Oct;7(10):743–746. doi: 10.1002/eji.1830071018. [DOI] [PubMed] [Google Scholar]
  22. Ishihara A., Hou Y., Jacobson K. The Thy-1 antigen exhibits rapid lateral diffusion in the plasma membrane of rodent lymphoid cells and fibroblasts. Proc Natl Acad Sci U S A. 1987 Mar;84(5):1290–1293. doi: 10.1073/pnas.84.5.1290. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Kammer G. M., Walter E. I., Medof M. E. Association of cytoskeletal re-organization with capping of the complement decay-accelerating factor on T lymphocytes. J Immunol. 1988 Nov 1;141(9):2924–2928. [PubMed] [Google Scholar]
  24. Kroczek R. A., Gunter K. C., Germain R. N., Shevach E. M. Thy-1 functions as a signal transduction molecule in T lymphocytes and transfected B lymphocytes. Nature. 1986 Jul 10;322(6075):181–184. doi: 10.1038/322181a0. [DOI] [PubMed] [Google Scholar]
  25. Kroczek R. A., Gunter K. C., Seligmann B., Shevach E. M. Induction of T cell activation by monoclonal anti-Thy-1 antibodies. J Immunol. 1986 Jun 15;136(12):4379–4384. [PubMed] [Google Scholar]
  26. Lazarovits J., Roth M. A single amino acid change in the cytoplasmic domain allows the influenza virus hemagglutinin to be endocytosed through coated pits. Cell. 1988 Jun 3;53(5):743–752. doi: 10.1016/0092-8674(88)90092-x. [DOI] [PubMed] [Google Scholar]
  27. Lennon V. A., Unger M., Dulbecco R. Thy-1: a differentiation marker of potential mammary myoepithelial cells in vitro. Proc Natl Acad Sci U S A. 1978 Dec;75(12):6093–6097. doi: 10.1073/pnas.75.12.6093. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Lesley J. F., Lennon V. A. Transitory expression of Thy-1 antigen in skeletal muscle development. Nature. 1977 Jul 14;268(5616):163–165. doi: 10.1038/268163a0. [DOI] [PubMed] [Google Scholar]
  29. Lesley J. F., Schulte R. J. Inhibition of cell growth by monoclonal anti-transferrin receptor antibodies. Mol Cell Biol. 1985 Aug;5(8):1814–1821. doi: 10.1128/mcb.5.8.1814. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Lisanti M. P., Sargiacomo M., Graeve L., Saltiel A. R., Rodriguez-Boulan E. Polarized apical distribution of glycosyl-phosphatidylinositol-anchored proteins in a renal epithelial cell line. Proc Natl Acad Sci U S A. 1988 Dec;85(24):9557–9561. doi: 10.1073/pnas.85.24.9557. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Low M. G., Saltiel A. R. Structural and functional roles of glycosyl-phosphatidylinositol in membranes. Science. 1988 Jan 15;239(4837):268–275. doi: 10.1126/science.3276003. [DOI] [PubMed] [Google Scholar]
  32. MacDonald H. R., Bron C., Rousseaux M., Horvath C., Cerottini J. C. Production and characterization of monoclonal anti-Thy-1 antibodies that stimulate lymphokine production by cytolytic T cell clones. Eur J Immunol. 1985 May;15(5):495–501. doi: 10.1002/eji.1830150514. [DOI] [PubMed] [Google Scholar]
  33. Machy P., Truneh A., Gennaro D., Hoffstein S. Major histocompatibility complex class I molecules internalized via coated pits in T lymphocytes. Nature. 1987 Aug 20;328(6132):724–726. doi: 10.1038/328724a0. [DOI] [PubMed] [Google Scholar]
  34. Malek T. R., Ortega G., Chan C., Kroczek R. A., Shevach E. M. Role of Ly-6 in lymphocyte activation. II. Induction of T cell activation by monoclonal anti-Ly-6 antibodies. J Exp Med. 1986 Sep 1;164(3):709–722. doi: 10.1084/jem.164.3.709. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Matsuura S., Eto S., Kato K., Tashiro Y. Ferritin immunoelectron microscopic localization of 5'-nucleotidase on rat liver cell surface. J Cell Biol. 1984 Jul;99(1 Pt 1):166–173. doi: 10.1083/jcb.99.1.166. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Morris R. Thy-1 in developing nervous tissue. Dev Neurosci. 1985;7(3):133–160. doi: 10.1159/000112283. [DOI] [PubMed] [Google Scholar]
  37. Mostov K. E., de Bruyn Kops A., Deitcher D. L. Deletion of the cytoplasmic domain of the polymeric immunoglobulin receptor prevents basolateral localization and endocytosis. Cell. 1986 Nov 7;47(3):359–364. doi: 10.1016/0092-8674(86)90592-1. [DOI] [PubMed] [Google Scholar]
  38. Murphy T. L., Decker G., August J. T. Glycoproteins of coated pits, cell junctions, and the entire cell surface revealed by monoclonal antibodies and immunoelectron microscopy. J Cell Biol. 1983 Aug;97(2):533–541. doi: 10.1083/jcb.97.2.533. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Oi V. T., Jones P. P., Goding J. W., Herzenberg L. A., Herzenberg L. A. Properties of monoclonal antibodies to mouse Ig allotypes, H-2, and Ia antigens. Curr Top Microbiol Immunol. 1978;81:115–120. doi: 10.1007/978-3-642-67448-8_18. [DOI] [PubMed] [Google Scholar]
  40. Parekh R. B., Tse A. G., Dwek R. A., Williams A. F., Rademacher T. W. Tissue-specific N-glycosylation, site-specific oligosaccharide patterns and lentil lectin recognition of rat Thy-1. EMBO J. 1987 May;6(5):1233–1244. doi: 10.1002/j.1460-2075.1987.tb02359.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Pearse B. M. Characterization of coated-vesicle adaptors: their reassembly with clathrin and with recycling receptors. Methods Cell Biol. 1989;31:229–246. doi: 10.1016/s0091-679x(08)61612-x. [DOI] [PubMed] [Google Scholar]
  42. Phelps B. M., Primakoff P., Koppel D. E., Low M. G., Myles D. G. Restricted lateral diffusion of PH-20, a PI-anchored sperm membrane protein. Science. 1988 Jun 24;240(4860):1780–1782. doi: 10.1126/science.3381102. [DOI] [PubMed] [Google Scholar]
  43. Pont S., Regnier-Vigouroux A., Naquet P., Blanc D., Pierres A., Marchetto S., Pierres M. Analysis of the Thy-1 pathway of T cell hybridoma activation using 17 rat monoclonal antibodies reactive with distinct Thy-1 epitopes. Eur J Immunol. 1985 Dec;15(12):1222–1228. doi: 10.1002/eji.1830151215. [DOI] [PubMed] [Google Scholar]
  44. Prywes R., Livneh E., Ullrich A., Schlessinger J. Mutations in the cytoplasmic domain of EGF receptor affect EGF binding and receptor internalization. EMBO J. 1986 Sep;5(9):2179–2190. doi: 10.1002/j.1460-2075.1986.tb04482.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Reichner J. S., Whiteheart S. W., Hart G. W. Intracellular trafficking of cell surface sialoglycoconjugates. J Biol Chem. 1988 Nov 5;263(31):16316–16326. [PubMed] [Google Scholar]
  46. Rodman J. S., Seidman L., Farquhar M. G. The membrane composition of coated pits, microvilli, endosomes, and lysosomes is distinctive in the rat kidney proximal tubule cell. J Cell Biol. 1986 Jan;102(1):77–87. doi: 10.1083/jcb.102.1.77. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Rothenberger S., Iacopetta B. J., Kühn L. C. Endocytosis of the transferrin receptor requires the cytoplasmic domain but not its phosphorylation site. Cell. 1987 May 8;49(3):423–431. doi: 10.1016/0092-8674(87)90295-9. [DOI] [PubMed] [Google Scholar]
  48. Saleh M., Lang R. J., Bartlett P. F. Thy-1-mediated regulation of a low-threshold transient calcium current in cultured sensory neurons. Proc Natl Acad Sci U S A. 1988 Jun;85(12):4543–4547. doi: 10.1073/pnas.85.12.4543. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Salisbury J. L., Condeelis J. S., Satir P. Role of coated vesicles, microfilaments, and calmodulin in receptor-mediated endocytosis by cultured B lymphoblastoid cells. J Cell Biol. 1980 Oct;87(1):132–141. doi: 10.1083/jcb.87.1.132. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Schmitt-Verhulst A. M., Guimezanes A., Boyer C., Poenie M., Tsien R., Buferne M., Hua C., Leserman L. Pleiotropic loss of activation pathways in a T-cell receptor alpha-chain deletion variant of a cytolytic T-cell clone. Nature. 1987 Feb 12;325(6105):628–631. doi: 10.1038/325628a0. [DOI] [PubMed] [Google Scholar]
  51. Stern P. L. Theta alloantigen on mouse and rat fibroblasts. Nat New Biol. 1973 Nov 21;246(151):76–78. doi: 10.1038/newbio246076a0. [DOI] [PubMed] [Google Scholar]
  52. Stroynowski I., Soloski M., Low M. G., Hood L. A single gene encodes soluble and membrane-bound forms of the major histocompatibility Qa-2 antigen: anchoring of the product by a phospholipid tail. Cell. 1987 Aug 28;50(5):759–768. doi: 10.1016/0092-8674(87)90334-5. [DOI] [PubMed] [Google Scholar]
  53. Tausk F., Fey M., Gigli I. Endocytosis and shedding of the decay accelerating factor on human polymorphonuclear cells. J Immunol. 1989 Nov 15;143(10):3295–3302. [PubMed] [Google Scholar]
  54. Thilo L. Quantification of endocytosis-derived membrane traffic. Biochim Biophys Acta. 1985 Sep 9;822(2):243–266. doi: 10.1016/0304-4157(85)90010-3. [DOI] [PubMed] [Google Scholar]
  55. Trowbridge I. S., Hyman R., Mazauskas C. The synthesis and properties of T25 blycoprotein in Thy-1-negative mutant lymphoma cells. Cell. 1978 May;14(1):21–32. doi: 10.1016/0092-8674(78)90297-0. [DOI] [PubMed] [Google Scholar]
  56. Tse D. B., Pernis B. Spontaneous internalization of Class I major histocompatibility complex molecules in T lymphoid cells. J Exp Med. 1984 Jan 1;159(1):193–207. doi: 10.1084/jem.159.1.193. [DOI] [PMC free article] [PubMed] [Google Scholar]
  57. Turner C. E., Newton M. R., Shotton D. M. Cytoskeletal involvement in the sequential capping of rat thymocyte surface glycoproteins. J Cell Sci. 1988 Mar;89(Pt 3):309–319. doi: 10.1242/jcs.89.3.309. [DOI] [PubMed] [Google Scholar]
  58. Turner J. R., Tartakoff A. M., Berger M. Intracellular degradation of the complement C3b/C4b receptor in the absence of ligand. J Biol Chem. 1988 Apr 5;263(10):4914–4920. [PubMed] [Google Scholar]
  59. Vega M. A., Strominger J. L. Constitutive endocytosis of HLA class I antigens requires a specific portion of the intracytoplasmic tail that shares structural features with other endocytosed molecules. Proc Natl Acad Sci U S A. 1989 Apr;86(8):2688–2692. doi: 10.1073/pnas.86.8.2688. [DOI] [PMC free article] [PubMed] [Google Scholar]
  60. Widnell C. C., Schneider Y. J., Pierre B., Baudhuin P., Trouet A. Evidence for a continual exchange of 5'-nucleotidase between the cell surface and cytoplasmic membranes in cultured rat fibroblasts. Cell. 1982 Jan;28(1):61–70. doi: 10.1016/0092-8674(82)90375-0. [DOI] [PubMed] [Google Scholar]
  61. Yeh E. T., Reiser H., Bamezai A., Rock K. L. TAP transcription and phosphatidylinositol linkage mutants are defective in activation through the T cell receptor. Cell. 1988 Mar 11;52(5):665–674. doi: 10.1016/0092-8674(88)90404-7. [DOI] [PubMed] [Google Scholar]
  62. Yeh E. T., Reiser H., Daley J., Rock K. L. Stimulation of T cells via the TAP molecule, a member in a family of activating proteins encoded in the Ly-6 locus. J Immunol. 1987 Jan 1;138(1):91–97. [PubMed] [Google Scholar]
  63. van den Bosch R. A., du Maine A. P., Geuze H. J., van der Ende A., Strous G. J. Recycling of 5'-nucleotidase in a rat hepatoma cell line. EMBO J. 1988 Nov;7(11):3345–3351. doi: 10.1002/j.1460-2075.1988.tb03206.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

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