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Isolation and ultrastructural characterization of squid synaptic vesicles

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dc.contributor.author Pekkurnaz, Gulcin
dc.contributor.author Fera, Andrea
dc.contributor.author Zimmerberg-Helms, Jessica
dc.contributor.author DeGiorgis, Joseph A.
dc.contributor.author Bezrukov, Ludmila
dc.contributor.author Blank, Paul S.
dc.contributor.author Mazar, Julia
dc.contributor.author Reese, Thomas S.
dc.contributor.author Zimmerberg, Joshua
dc.date.accessioned 2011-05-09T19:15:51Z
dc.date.available 2011-05-09T19:15:51Z
dc.date.issued 2011-04
dc.identifier.citation Biological Bulletin 220 (2011): 89-96 en_US
dc.identifier.uri http://hdl.handle.net/1912/4555
dc.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. en_US
dc.description.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. en_US
dc.description.sponsorship JAD is supported by the RI-INBRE program award # P20RR016457-10 from the National Center for Research Resources (NCRR), NIH. en_US
dc.format.mimetype application/pdf
dc.language.iso en_US en_US
dc.publisher Marine Biological Laboratory en_US
dc.relation.uri http://www.biolbull.org/cgi/content/abstract/220/2/89?ct=ct
dc.title Isolation and ultrastructural characterization of squid synaptic vesicles en_US
dc.type Article en_US


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