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PMID: 7698987 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Cellubrevin and synaptobrevins: similar subcellular localization and biochemical properties in PC12 cells.

The Journal of cell biology ·Vol. 129 ·No. 1 ·1995-04-00 ·Pages 219-31

Chilcote TJ, Galli T, Mundigl O, Edelmann L, McPherson PS, Takei K, De Camilli P

Abstract

There is strong evidence to indicate that proteins of the synaptobrevin family play a key role in exocytosis. Synaptobrevin 1 and 2 are expressed at high concentration in brain where they are localized on synaptic vesicles. Cellubrevin, a very similar protein, has a widespread tissue distribution and in fibroblasts is localized on endosome-derived, transferin receptor-positive vesicles. Since brain cellubrevin is not detectable in synaptic vesicles, we investigated whether cellubrevin and the synaptobrevins are differentially targeted when co-expressed in the same cell. We report that in the nervous system cellubrevin is expressed at significant levels only by glia and vascular cells. However, cellubrevin is coexpressed with the two synaptobrevins in PC12 cells, a neuroendocrine cell line which contains synaptic vesicle-like microvesicles. In PC12 cells, cellubrevin has a distribution very similar to that of synaptobrevin 1 and 2. The three proteins are targeted to neurites which exclude the transferrin receptor and are enriched in synaptic-like microvesicles and dense-core granules. They are recovered in the synaptic-like microvesicle peak of glycerol velocity gradients, have a similar distribution in isopycnic fractionation and are coprecipitated by anti-synaptobrevin 2 immunobeads. Finally, cellubrevin, like the synaptobrevins, interact with the neuronal t-SNAREs syntaxin 1 and SNAP-25. These results suggest that cellubrevin and the synaptobrevins have similar function and do not play a specialized role in constitutive and regulated exocytosis, respectively.

MeSH Terms
Amino Acid Sequence Animals Antibodies Brain/metabolism Cell Fractionation Cells, Cultured Centrifugation, Density Gradient Cytoplasmic Granules/metabolism,ultrastructure Fetus Hippocampus/metabolism Membrane Proteins/analysis,biosynthesis,isolation & purification,metabolism Molecular Sequence Data Nerve Tissue Proteins/analysis,biosynthesis,isolation & purification,metabolism Neurites/metabolism,ultrastructure Neurons/metabolism Organelles/metabolism,ultrastructure PC12 Cells Qa-SNARE Proteins R-SNARE Proteins Rats Synapses/metabolism,ultrastructure Synaptosomal-Associated Protein 25 Syntaxin 1 Vesicle-Associated Membrane Protein 3
Chemicals
Antibodies Membrane Proteins Nerve Tissue Proteins Qa-SNARE Proteins R-SNARE Proteins Snap25 protein, rat Stx1a protein, rat Synaptosomal-Associated Protein 25 Syntaxin 1 Vesicle-Associated Membrane Protein 3
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Chilcote T J
Department of Cell Biology, Yale University School of Medicine, New Haven, Connecticut 06510.
Galli T
Mundigl O
Edelmann L
McPherson P S
Takei K
De Camilli P
References (64)
64 references, click to expand
  1. GABA and pancreatic beta-cells: colocalization of glutamic acid decarboxylase (GAD) and GABA with synaptic-like microvesicles suggests their role in GABA storage and secretion.
    EMBO J. 1991 May;10(5):1275-84 PMID: 2022191
  2. Synaptobrevin binding to synaptophysin: a potential mechanism for controlling the exocytotic fusion machine.
    EMBO J. 1995 Jan 16;14(2):224-31 PMID: 7835333
  3. Synaptotagmin II. A novel differentially distributed form of synaptotagmin.
    J Biol Chem. 1991 Jul 25;266(21):13548-52 PMID: 1856191
  4. Synaptophysin is sorted from endocytotic markers in neuroendocrine PC12 cells but not transfected fibroblasts.
    Neuron. 1991 Aug;7(2):309-17 PMID: 1908253
  5. Colocalization of synaptophysin with transferrin receptors: implications for synaptic vesicle biogenesis.
    J Cell Biol. 1991 Oct;115(1):151-64 PMID: 1918133
  6. Newly synthesized synaptophysin is transported to synaptic-like microvesicles via constitutive secretory vesicles and the plasma membrane.
    EMBO J. 1991 Dec;10(12):3589-601 PMID: 1935891
  7. A family of proteins involved in intracellular transport.
    Cell. 1992 Jan 24;68(2):181-2 PMID: 1733497
  8. Ca2+ stores in Purkinje neurons: endoplasmic reticulum subcompartments demonstrated by the heterogeneous distribution of the InsP3 receptor, Ca(2+)-ATPase, and calsequestrin.
    J Neurosci. 1992 Feb;12(2):489-505 PMID: 1311032
  9. Exo-endocytotic recycling of synaptic vesicles in developing processes of cultured hippocampal neurons.
    J Cell Biol. 1992 May;117(4):849-61 PMID: 1577861
  10. Members of the VAMP family of synaptic vesicle proteins are components of glucose transporter-containing vesicles from rat adipocytes.
    J Biol Chem. 1992 Jun 15;267(17):11681-4 PMID: 1601842
  11. Clathrin-coated vesicles in nervous tissue are involved primarily in synaptic vesicle recycling.
    J Cell Biol. 1992 Sep;118(6):1379-88 PMID: 1325974
  12. Tetanus and botulinum-B neurotoxins block neurotransmitter release by proteolytic cleavage of synaptobrevin.
    Nature. 1992 Oct 29;359(6398):832-5 PMID: 1331807
  13. Movement from trans-Golgi network to cell surface in semiintact cells.
    Methods Enzymol. 1992;219:234-48 PMID: 1487997
  14. SNAP receptors implicated in vesicle targeting and fusion.
    Nature. 1993 Mar 25;362(6418):318-24 PMID: 8455717
  15. Brain contains two forms of synaptic vesicle protein 2.
    Proc Natl Acad Sci U S A. 1993 Mar 15;90(6):2150-4 PMID: 7681585
  16. The molecular machinery for secretion is conserved from yeast to neurons.
    Proc Natl Acad Sci U S A. 1993 Apr 1;90(7):2559-63 PMID: 8096639
  17. Comparison of the intracellular effects of clostridial neurotoxins on exocytosis from streptolysin O-permeabilized rat pheochromocytoma (PC 12) and bovine adrenal chromaffin cells.
    Neuroscience. 1993 Mar;53(2):547-52 PMID: 8492915
  18. A gamma-aminobutyric acid transporter driven by a proton pump is present in synaptic-like microvesicles of pancreatic beta cells.
    Proc Natl Acad Sci U S A. 1993 Jun 1;90(11):5317-21 PMID: 8506380
  19. Cellubrevin is a ubiquitous tetanus-toxin substrate homologous to a putative synaptic vesicle fusion protein.
    Nature. 1993 Jul 22;364(6435):346-9 PMID: 8332193
  20. Selective storage of acetylcholine, but not catecholamines, in neuroendocrine synaptic-like microvesicles of early endosomal origin.
    Neuron. 1993 Jul;11(1):105-21 PMID: 8338662
  21. Calculation and control of free divalent cations in solutions used for membrane fusion studies.
    Methods Enzymol. 1993;221:149-57 PMID: 8361372
  22. Synaptotagmin: a membrane constituent of neuropeptide-containing large dense-core vesicles.
    J Neurosci. 1993 Sep;13(9):3895-903 PMID: 8366350
  23. Botulinum neurotoxin A selectively cleaves the synaptic protein SNAP-25.
    Nature. 1993 Sep 9;365(6442):160-3 PMID: 8103915
  24. Homologs of the synaptobrevin/VAMP family of synaptic vesicle proteins function on the late secretory pathway in S. cerevisiae.
    Cell. 1993 Sep 10;74(5):855-61 PMID: 8374953
  25. The syntaxin family of vesicular transport receptors.
    Cell. 1993 Sep 10;74(5):863-73 PMID: 7690687
  26. Synaptic vesicle proteins and early endosomes in cultured hippocampal neurons: differential effects of Brefeldin A in axon and dendrites.
    J Cell Biol. 1993 Sep;122(6):1207-21 PMID: 8376458
  27. Differential expression of transcripts from syb, a Drosophila melanogaster gene encoding VAMP (synaptobrevin) that is abundant in non-neuronal cells.
    Gene. 1993 Sep 15;131(2):175-81 PMID: 8406010
  28. A protein assembly-disassembly pathway in vitro that may correspond to sequential steps of synaptic vesicle docking, activation, and fusion.
    Cell. 1993 Nov 5;75(3):409-18 PMID: 8221884
  29. Botulinum neurotoxin C1 blocks neurotransmitter release by means of cleaving HPC-1/syntaxin.
    EMBO J. 1993 Dec;12(12):4821-8 PMID: 7901002
  30. Identification of the nerve terminal targets of botulinum neurotoxin serotypes A, D, and E.
    J Biol Chem. 1993 Nov 15;268(32):23784-7 PMID: 8226912
  31. Identification and characterization of Drosophila genes for synaptic vesicle proteins.
    J Neurosci. 1993 Nov;13(11):4924-35 PMID: 8229205
  32. Relative properties and localizations of synaptic vesicle protein isoforms: the case of the synaptophysins.
    J Neurosci. 1993 Nov;13(11):4997-5007 PMID: 8229211
  33. Cleavage of members of the synaptobrevin/VAMP family by types D and F botulinal neurotoxins and tetanus toxin.
    J Biol Chem. 1994 Apr 29;269(17):12764-72 PMID: 8175689
  34. VAMP-1: a synaptic vesicle-associated integral membrane protein.
    Proc Natl Acad Sci U S A. 1988 Jun;85(12):4538-42 PMID: 3380805
  35. Fractionation of synaptophysin-containing vesicles from rat brain and cultured PC12 pheochromocytoma cells.
    FEBS Lett. 1988 Nov 21;240(1-2):71-7 PMID: 3142795
  36. The tetanus toxin light chain inhibits exocytosis.
    FEBS Lett. 1989 Jan 2;242(2):245-8 PMID: 2914606
  37. Synaptobrevin: an integral membrane protein of 18,000 daltons present in small synaptic vesicles of rat brain.
    EMBO J. 1989 Feb;8(2):379-84 PMID: 2498078
  38. Two vesicle-associated membrane protein genes are differentially expressed in the rat central nervous system.
    J Biol Chem. 1989 Jul 5;264(19):11061-4 PMID: 2472388
  39. Spatial segregation of the regulated and constitutive secretory pathways.
    J Cell Biol. 1989 Jul;109(1):51-60 PMID: 2545730
  40. Poration by alpha-toxin and streptolysin O: an approach to analyze intracellular processes.
    Methods Cell Biol. 1989;31:63-90 PMID: 2779453
  41. Growth and cultivation of dissociated neurons and glial cells from embryonic chick, rat and human brain in flask cultures.
    Neurobiology. 1972;2(3):97-105 PMID: 4572654
  42. Establishment of a noradrenergic clonal line of rat adrenal pheochromocytoma cells which respond to nerve growth factor.
    Proc Natl Acad Sci U S A. 1976 Jul;73(7):2424-8 PMID: 1065897
  43. Rat hippocampal neurons in dispersed cell culture.
    Brain Res. 1977 May 13;126(3):397-42 PMID: 861729
  44. A simplification of the protein assay method of Lowry et al. which is more generally applicable.
    Anal Biochem. 1977 Dec;83(2):346-56 PMID: 603028
  45. Morphologic and cytochemical properties of a clonal line of rat adrenal pheochromocytoma cells which respond to nerve growth factor.
    Lab Invest. 1978 Aug;39(2):77-89 PMID: 682602
  46. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications.
    Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350-4 PMID: 388439
  47. Continuous, clonal, insulin- and somatostatin-secreting cell lines established from a transplantable rat islet cell tumor.
    Proc Natl Acad Sci U S A. 1980 Jun;77(6):3519-23 PMID: 6106192
  48. Calcium-evoked secretion from digitonin-permeabilized adrenal medullary chromaffin cells.
    J Biol Chem. 1983 Apr 25;258(8):4994-5000 PMID: 6300108
  49. 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):1337-54 PMID: 6404910
  50. An electron microscopic study of the development of axons and dendrites by hippocampal neurons in culture. I. Cells which develop without intercellular contacts.
    J Neurosci. 1984 Aug;4(8):1944-53 PMID: 6470762
  51. Secretogranins I and II: two tyrosine-sulfated secretory proteins common to a variety of cells secreting peptides by the regulated pathway.
    J Cell Biol. 1985 Nov;101(5 Pt 1):1999-2011 PMID: 4055903
  52. Protein p38: an integral membrane protein specific for small vesicles of neurons and neuroendocrine cells.
    J Cell Biol. 1986 Dec;103(6 Pt 1):2511-27 PMID: 3097029
  53. Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa.
    Anal Biochem. 1987 Nov 1;166(2):368-79 PMID: 2449095
  54. Synapsins: mosaics of shared and individual domains in a family of synaptic vesicle phosphoproteins.
    Science. 1989 Sep 29;245(4925):1474-80 PMID: 2506642
  55. Microvesicles of the neurohypophysis are biochemically related to small synaptic vesicles of presynaptic nerve terminals.
    J Cell Biol. 1989 Dec;109(6 Pt 2):3425-33 PMID: 2513331
  56. Distinct patterns of expression of two VAMP genes within the rat brain.
    J Neurosci. 1990 Apr;10(4):1380-7 PMID: 2329380
  57. Polarized sorting of viral glycoproteins to the axon and dendrites of hippocampal neurons in culture.
    Cell. 1990 Jul 13;62(1):63-72 PMID: 2163770
  58. Synaptic vesicles immunoisolated from rat cerebral cortex contain high levels of glutamate.
    Neuron. 1989 Dec;3(6):715-20 PMID: 2577130
  59. Expression of the synaptic vesicle proteins VAMPs/synaptobrevins 1 and 2 in non-neural tissues.
    J Biol Chem. 1994 Jun 3;269(22):15403-6 PMID: 8195178
  60. Tetanus toxin-mediated cleavage of cellubrevin impairs exocytosis of transferrin receptor-containing vesicles in CHO cells.
    J Cell Biol. 1994 Jun;125(5):1015-24 PMID: 8195285
  61. Tetanus toxin light chain cleaves a vesicle-associated membrane protein (VAMP) isoform 2 in rat pancreatic zymogen granules and inhibits enzyme secretion.
    J Biol Chem. 1994 Jun 24;269(25):17062-6 PMID: 7516331
  62. Vesicle fusion from yeast to man.
    Nature. 1994 Jul 21;370(6486):191-3 PMID: 8028665
  63. p145, a major Grb2-binding protein in brain, is co-localized with dynamin in nerve terminals where it undergoes activity-dependent dephosphorylation.
    J Biol Chem. 1994 Dec 2;269(48):30132-9 PMID: 7982917
  64. Developmental expression of the 25-kDa synaptosomal-associated protein (SNAP-25) in rat brain.
    Proc Natl Acad Sci U S A. 1991 Jun 15;88(12):5247-51 PMID: 1711221
Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.129.1.219
Published
1995-04-00
Pages
219-31
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2120369
Subset
IM
Grants
NCI NIH HHS · P01-CA46128 · United States
External Links
PubMed source DOI: 10.1083/jcb.129.1.219 PMC Full Text View journal details
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