Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1985 Apr 1;100(4):1284–1294. doi: 10.1083/jcb.100.4.1284

Identification of a transmembrane glycoprotein specific for secretory vesicles of neural and endocrine cells

PMCID: PMC2113776  PMID: 2579958

Abstract

Several types of cells store proteins in secretory vesicles from which they are released by an appropriate stimulus. It might be expected that the secretory vesicles in different cell types use similar molecular machinery. Here we describe a transmembrane glycoprotein (Mr approximately 100,000) that is present in secretory vesicles in all neurons and endocrine cells studied, in species from elasmobranch fish to mammals, and in neural and endocrine cell lines. It was detected by cross-reactivity with monoclonal antibodies raised to highly purified cholinergic synaptic vesicles from the electric organ of fish. By immunoprecipitation of intact synaptic vesicles and electron microscopic immunoperoxidase labeling, we have shown that the antigenic determinant is on the cytoplasmic face of the synaptic vesicles. However, the electrophoretic mobility of the antigen synthesized in the presence of tunicamycin is reduced to Mr approximately 62,000, which suggests that the antigen is glycosylated and must therefore span the vesicle membrane.

Full Text

The Full Text of this article is available as a PDF (4.2 MB).

Selected References

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

  1. Bishop A. E., Polak J. M., Facer P., Ferri G. L., Marangos P. J., Pearse A. G. Neuron specific enolase: a common marker for the endocrine cells and innervation of the gut and pancreas. Gastroenterology. 1982 Oct;83(4):902–915. [PubMed] [Google Scholar]
  2. Bordier C. Phase separation of integral membrane proteins in Triton X-114 solution. J Biol Chem. 1981 Feb 25;256(4):1604–1607. [PubMed] [Google Scholar]
  3. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  4. Buckley K. M., Schweitzer E. S., Miljanich G. P., Clift-O'Grady L., Kushner P. D., Reichardt L. F., Kelly R. B. A synaptic vesicle antigen is restricted to the junctional region of the presynaptic plasma membrane. Proc Natl Acad Sci U S A. 1983 Dec;80(23):7342–7346. doi: 10.1073/pnas.80.23.7342. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Burke B., Griffiths G., Reggio H., Louvard D., Warren G. A monoclonal antibody against a 135-K Golgi membrane protein. EMBO J. 1982;1(12):1621–1628. doi: 10.1002/j.1460-2075.1982.tb01364.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Burnette W. N. "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal Biochem. 1981 Apr;112(2):195–203. doi: 10.1016/0003-2697(81)90281-5. [DOI] [PubMed] [Google Scholar]
  7. Carlson S. S., Kelly R. B. A highly antigenic proteoglycan-like component of cholinergic synaptic vesicles. J Biol Chem. 1983 Sep 25;258(18):11082–11091. [PubMed] [Google Scholar]
  8. Carlson S. S., Kelly R. B. An antiserum specific for cholinergic synaptic vesicles from electric organ. J Cell Biol. 1980 Oct;87(1):98–103. doi: 10.1083/jcb.87.1.98. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Carlson S. S., Wagner J. A., Kelly R. B. Purification of synaptic vesicles from elasmobranch electric organ and the use of biophysical criteria to demonstrate purity. Biochemistry. 1978 Apr 4;17(7):1188–1199. doi: 10.1021/bi00600a009. [DOI] [PubMed] [Google Scholar]
  10. De Camilli P., Cameron R., Greengard P. Synapsin I (protein I), a nerve terminal-specific phosphoprotein. I. Its general distribution in synapses of the central and peripheral nervous system demonstrated by immunofluorescence in frozen and plastic sections. J Cell Biol. 1983 May;96(5):1337–1354. doi: 10.1083/jcb.96.5.1337. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. GRAY E. G., WHITTAKER V. P. The isolation of nerve endings from brain: an electron-microscopic study of cell fragments derived by homogenization and centrifugation. J Anat. 1962 Jan;96:79–88. [PMC free article] [PubMed] [Google Scholar]
  12. Green R., Shields D. Somatostatin discriminates between the intracellular pathways of secretory and membrane proteins. J Cell Biol. 1984 Jul;99(1 Pt 1):97–104. doi: 10.1083/jcb.99.1.97. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gumbiner B., Kelly R. B. Secretory granules of an anterior pituitary cell line, AtT-20, contain only mature forms of corticotropin and beta-lipotropin. Proc Natl Acad Sci U S A. 1981 Jan;78(1):318–322. doi: 10.1073/pnas.78.1.318. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Gumbiner B., Kelly R. B. Two distinct intracellular pathways transport secretory and membrane glycoproteins to the surface of pituitary tumor cells. Cell. 1982 Jan;28(1):51–59. doi: 10.1016/0092-8674(82)90374-9. [DOI] [PubMed] [Google Scholar]
  15. Hoffman S., Sorkin B. C., White P. C., Brackenbury R., Mailhammer R., Rutishauser U., Cunningham B. A., Edelman G. M. Chemical characterization of a neural cell adhesion molecule purified from embryonic brain membranes. J Biol Chem. 1982 Jul 10;257(13):7720–7729. [PubMed] [Google Scholar]
  16. Hooper J. E., Carlson S. S., Kelly R. B. Antibodies to synaptic vesicles purified from Narcine electric organ bind a subclass of mammalian nerve terminals. J Cell Biol. 1980 Oct;87(1):104–113. doi: 10.1083/jcb.87.1.104. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hubbard S. C., Ivatt R. J. Synthesis and processing of asparagine-linked oligosaccharides. Annu Rev Biochem. 1981;50:555–583. doi: 10.1146/annurev.bi.50.070181.003011. [DOI] [PubMed] [Google Scholar]
  18. Johnson G. D., Davidson R. S., McNamee K. C., Russell G., Goodwin D., Holborow E. J. Fading of immunofluorescence during microscopy: a study of the phenomenon and its remedy. J Immunol Methods. 1982 Dec 17;55(2):231–242. doi: 10.1016/0022-1759(82)90035-7. [DOI] [PubMed] [Google Scholar]
  19. Kelly R. B., Buckley K. M., Burgess T. L., Carlson S. S., Caroni P., Hooper J. E., Katzen A., Moore H. P., Pfeffer S. R., Schroer T. A. Membrane traffic in neurons and peptide-secreting cells. Cold Spring Harb Symp Quant Biol. 1983;48(Pt 2):697–705. doi: 10.1101/sqb.1983.048.01.073. [DOI] [PubMed] [Google Scholar]
  20. Krieger D. T. Brain peptides: what, where, and why? Science. 1983 Dec 2;222(4627):975–985. doi: 10.1126/science.6139875. [DOI] [PubMed] [Google Scholar]
  21. Köhler G., Milstein C. Derivation of specific antibody-producing tissue culture and tumor lines by cell fusion. Eur J Immunol. 1976 Jul;6(7):511–519. doi: 10.1002/eji.1830060713. [DOI] [PubMed] [Google Scholar]
  22. Lander A. D., Fujii D. K., Gospodarowicz D., Reichardt L. F. Characterization of a factor that promotes neurite outgrowth: evidence linking activity to a heparan sulfate proteoglycan. J Cell Biol. 1982 Sep;94(3):574–585. doi: 10.1083/jcb.94.3.574. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Le Roith D., Shiloach J., Roth J. Is there an earlier phylogenetic precursor that is common to both the nervous and endocrine systems? Peptides. 1982 May-Jun;3(3):211–215. doi: 10.1016/0196-9781(82)90080-8. [DOI] [PubMed] [Google Scholar]
  24. Lin J. J., Queally S. A. A monoclonal antibody that recognizes Golgi-associated protein of cultured fibroblast cells. J Cell Biol. 1982 Jan;92(1):108–112. doi: 10.1083/jcb.92.1.108. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Louvard D., Morris C., Warren G., Stanley K., Winkler F., Reggio H. A monoclonal antibody to the heavy chain of clathrin. EMBO J. 1983;2(10):1655–1664. doi: 10.1002/j.1460-2075.1983.tb01640.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Louvard D., Reggio H., Warren G. Antibodies to the Golgi complex and the rough endoplasmic reticulum. J Cell Biol. 1982 Jan;92(1):92–107. doi: 10.1083/jcb.92.1.92. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Mains R. E., Eipper B. A. Biosynthesis of adrenocorticotropic hormone in mouse pituitary tumor cells. J Biol Chem. 1976 Jul 10;251(13):4115–4120. [PubMed] [Google Scholar]
  28. Matthew W. D., Tsavaler L., Reichardt L. F. Identification of a synaptic vesicle-specific membrane protein with a wide distribution in neuronal and neurosecretory tissue. J Cell Biol. 1981 Oct;91(1):257–269. doi: 10.1083/jcb.91.1.257. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Moore H. P., Gumbiner B., Kelly R. B. A subclass of proteins and sulfated macromolecules secreted by AtT-20 (mouse pituitary tumor) cells is sorted with adrenocorticotropin into dense secretory granules. J Cell Biol. 1983 Sep;97(3):810–817. doi: 10.1083/jcb.97.3.810. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Roberts J. L., Phillips M., Rosa P. A., Herbert E. Steps involved in the processing of common precursor forms of adrenocorticotropin and endorphin in cultures of mouse pituitary cells. Biochemistry. 1978 Aug 22;17(17):3609–3618. doi: 10.1021/bi00610a030. [DOI] [PubMed] [Google Scholar]
  31. Rose J. K., Bergmann J. E. Expression from cloned cDNA of cell-surface secreted forms of the glycoprotein of vesicular stomatitis virus in eucaryotic cells. Cell. 1982 Oct;30(3):753–762. doi: 10.1016/0092-8674(82)90280-x. [DOI] [PubMed] [Google Scholar]
  32. Schmechel D., Marangos P. J., Brightman M. Neurone-specific enolase is a molecular marker for peripheral and central neuroendocrine cells. Nature. 1978 Dec 21;276(5690):834–836. doi: 10.1038/276834a0. [DOI] [PubMed] [Google Scholar]
  33. Smith Z. D., D'Eugenio-Gumkowski F., Yanagisawa K., Jamieson J. D. Endogenous and monoclonal antibodies to the rat pancreatic acinar cell Golgi complex. J Cell Biol. 1984 Jun;98(6):2035–2046. doi: 10.1083/jcb.98.6.2035. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Tartakoff A., Vassalli P., Détraz M. Comparative studies of intracellular transport of secretory proteins. J Cell Biol. 1978 Dec;79(3):694–707. doi: 10.1083/jcb.79.3.694. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Tougard C., Louvard D., Picart R., Tixier-Vidal A. The rough endoplasmic reticulum and the Golgi apparatus visualized using specific antibodies in normal and tumoral prolactin cells in culture. J Cell Biol. 1983 May;96(5):1197–1207. doi: 10.1083/jcb.96.5.1197. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Trifaró J. M., Dworkind J. A new and simple method for isolation of adrenal chromaffin granules by means of an isotonic density gradient. Anal Biochem. 1970 Apr;34(2):403–412. doi: 10.1016/0003-2697(70)90125-9. [DOI] [PubMed] [Google Scholar]
  37. de StGroth S. F., Scheidegger D. Production of monoclonal antibodies: strategy and tactics. J Immunol Methods. 1980;35(1-2):1–21. doi: 10.1016/0022-1759(80)90146-5. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES