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    Isolation and ultrastructural characterization of squid synaptic vesicles

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    Date
    2011-04
    Author
    Pekkurnaz, Gulcin  Concept link
    Fera, Andrea  Concept link
    Zimmerberg-Helms, Jessica  Concept link
    DeGiorgis, Joseph A.  Concept link
    Bezrukov, Ludmila  Concept link
    Blank, Paul S.  Concept link
    Mazar, Julia  Concept link
    Reese, Thomas S.  Concept link
    Zimmerberg, Joshua  Concept link
    Metadata
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    Citable URI
    https://hdl.handle.net/1912/4555
    As published
    https://doi.org/10.1086/BBLv220n2p89
    DOI
    10.1086/BBLv220n2p89
    Abstract
    Synaptic vesicles contain a variety of proteins and lipids that mediate fusion with the pre-synaptic membrane. Although the structures of many synaptic vesicle proteins are known, an overall picture of how they are organized at the vesicle surface is lacking. In this paper, we describe a better method for the isolation of squid synaptic vesicles and characterize the results. For highly pure and intact synaptic vesicles from squid optic lobe, glycerol density gradient centrifugation was the key step. Different electron microscopic methods show that vesicle membrane surfaces are largely covered with structures corresponding to surface proteins. Each vesicle contains several stalked globular structures that extend from the vesicle surface and are consistent with the V-ATPase. BLAST search of a library of squid expressed sequence tags identifies 10 V-ATPase subunits, which are expressed in the squid stellate ganglia. Negative-stain tomography demonstrates directly that vesicles flatten during the drying step of negative staining, and furthermore shows details of individual vesicles and other proteins at the vesicle surface.
    Description
    Author Posting. © Marine Biological Laboratory, 2011. This article is posted here by permission of Marine Biological Laboratory for personal use, not for redistribution. The definitive version was published in Biological Bulletin 220 (2011): 89-96.
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    Suggested Citation
    Biological Bulletin 220 (2011): 89-96
     
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