Fusion of green fluorescent protein to the C-terminus of granulysin alters its intracellular localization in comparison to the native molecule

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Fusion of green fluorescent protein to the C-terminus of granulysin alters its intracellular localization in comparison to the native molecule

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dc.contributor.author Hanson, Dennis A. en_US
dc.contributor.author Ziegler, Steven F. en_US
dc.date.accessioned 2010-04-21T15:53:36Z
dc.date.available 2010-04-21T15:53:36Z
dc.date.issued 2004 en_US
dc.identifier.citation Hanson D, Ziegler S. Fusion of green fluorescent protein to the C-terminus of granulysin alters its intracellular localization in comparison to the native molecule. Journal of Negative Results in BioMedicine. 2004;3(1):2. en_US
dc.identifier.other 10.1186/1477-5751-3-2 en_US
dc.identifier.uri http://www.jnrbm.com/content/3/1/2 en_US
dc.identifier.uri http://hdl.handle.net/1773/15755
dc.description.abstract The engineering of green fluorescent protein (GFP) fusion constructs in order to visibly tag a protein of interest has become a commonly used cell biology technique. Although caveats to this approach are obvious, literature reports in which the chimeric molecule behaves differently than the native molecule are scant. This brief report describes one such case. Granulysin, a small lytic and antimicrobial protein produced by cytotoxic lymphocytes, traffics to the regulated secretory system and is subsequently released from cells upon proper stimulus. In an attempt to elucidate mechanisms by which it accumulates in and is released from cytolytic granules, GFP was fused to the C-terminus of granulysin and expressed in an NK cell line. A control construct expressing the native protein was similarly expressed. The data demonstrate that, while the fusion protein is expressed and secreted, its subcellular localization is altered in comparison to native granulysin. Thus, the addition of GFP to the C-terminus of granulysin obscures the signal(s) that cytotoxic lymphocytes use to sort it to the regulated secretory pathway despite its normal biosynthesis and secretion. This example is offered as a cautionary account for other researchers who contemplate using this technology. en_US
dc.description.sponsorship en_US
dc.language.iso en_US en_US
dc.title Fusion of green fluorescent protein to the C-terminus of granulysin alters its intracellular localization in comparison to the native molecule en_US
dc.type Article en_US


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