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. 1979 Aug;76(8):4126–4130. doi: 10.1073/pnas.76.8.4126

Topological organization of proteins in an intracellular secretory organelle: the synaptic vesicle.

J A Wagner, R B Kelly
PMCID: PMC383991  PMID: 291069

Abstract

Intact synaptic vesicles prepared from the electric organ of the marine elasmobranch Narcine brasiliensis have eight major polypeptides demonstrable on sodium dodecyl sulfate gels. Six of these copurify with the synaptic vesicles during isolation of vesicles by chromatography on CPG-3000 and, by this criterion, are specific to vesicles. The other two are either shared by many membrane or are contaminants. One of these proteins comigrates with actin. Three different approaches were used to determine which proteins were exposed on the external, cytoplasmic surface of the vesicle and which were internal. The first was susceptibility to the proteases trypsin, Streptomyces griseus protease, and Pronase; the second was labeling by the membrane-impermeable reagent diazotized [125I]iodosulfanilic acid; and the third was iodination catalyzed by lactoperoxidase. In general, the three approaches give the same result: six of the eight proteins are on the external, cytoplasmic surface and two are accessible only after the vesicles are lysed by freezing and thawing or by detergents. Five of the vesicle-specific proteins are external and one is internal. The actin-like protein is internal. Proteins involved in the interaction of vesicles with the presynaptic membrane during exocytosis might be expected to be vesicle specific and external.

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Selected References

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

  1. Berl S., Puszkin S., Nicklas W. J. Actomyosin-like protein in brain. Science. 1973 Feb 2;179(4072):441–446. doi: 10.1126/science.179.4072.441. [DOI] [PubMed] [Google Scholar]
  2. 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]
  3. Dodge F. A., Jr, Rahamimoff R. Co-operative action a calcium ions in transmitter release at the neuromuscular junction. J Physiol. 1967 Nov;193(2):419–432. doi: 10.1113/jphysiol.1967.sp008367. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Fairbanks G., Jr, Levinthal C., Reeder R. H. Analysis of C14-labeled proteins by disc electrophoresis. Biochem Biophys Res Commun. 1965 Aug 16;20(4):393–399. doi: 10.1016/0006-291x(65)90589-9. [DOI] [PubMed] [Google Scholar]
  5. Heuser J. E., Reese T. S., Dennis M. J., Jan Y., Jan L., Evans L. Synaptic vesicle exocytosis captured by quick freezing and correlated with quantal transmitter release. J Cell Biol. 1979 May;81(2):275–300. doi: 10.1083/jcb.81.2.275. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Kelly R. B., Deutsch J. W., Carlson S. S., Wagner J. A. Biochemistry of neurotransmitter release. Annu Rev Neurosci. 1979;2:399–446. doi: 10.1146/annurev.ne.02.030179.002151. [DOI] [PubMed] [Google Scholar]
  7. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  8. Llinás R. R. Depolarization-release coupling systems in neurons. Neurosci Res Program Bull. 1977 Dec;15(4):555–687. [PubMed] [Google Scholar]
  9. Nagy A., Baker R. R., Morris S. J., Whittaker V. P. The preparation and characterization of synaptic vesicles of high purity. Brain Res. 1976 Jun 11;109(2):285–309. doi: 10.1016/0006-8993(76)90531-x. [DOI] [PubMed] [Google Scholar]
  10. Rothman J. E., Lenard J. Membrane asymmetry. Science. 1977 Feb 25;195(4280):743–753. doi: 10.1126/science.402030. [DOI] [PubMed] [Google Scholar]
  11. Schaffner W., Weissmann C. A rapid, sensitive, and specific method for the determination of protein in dilute solution. Anal Biochem. 1973 Dec;56(2):502–514. doi: 10.1016/0003-2697(73)90217-0. [DOI] [PubMed] [Google Scholar]
  12. Sears D. A., Reed C. F., Helmkamp R. W. A radioactive label for the erythrocyte membrane. Biochim Biophys Acta. 1971 Jun 1;233(3):716–719. doi: 10.1016/0005-2736(71)90170-2. [DOI] [PubMed] [Google Scholar]
  13. Stanley P. E., Williams S. G. Use of the liquid scintillation spectrometer for determining adenosine triphosphate by the luciferase enzyme. Anal Biochem. 1969 Jun;29(3):381–392. doi: 10.1016/0003-2697(69)90323-6. [DOI] [PubMed] [Google Scholar]
  14. Tashiro T., Stadler H. Chemical composition of cholinergic synaptic vesicles from Torpedo marmorata based on improved purification. Eur J Biochem. 1978 Oct 16;90(3):479–487. doi: 10.1111/j.1432-1033.1978.tb12627.x. [DOI] [PubMed] [Google Scholar]
  15. Wagner J. A., Carlson S. S., Kelly R. B. Chemical and physical characterization of cholinergic synaptic vesicles. Biochemistry. 1978 Apr 4;17(7):1199–1206. doi: 10.1021/bi00600a010. [DOI] [PubMed] [Google Scholar]

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