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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
4
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pubmed:dateCreated |
2008-10-6
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pubmed:abstractText |
The glutathione S-transferase (GST)-fused protein expression system has been extensively used to generate a large quantity of proteins and has served for functional analysis in vitro. In this study, we developed a novel approach for the efficient intracellular delivery of GST-fused proteins into living cells to expand their usefulness up to in vivo use. Since protein cationization techniques are powerful strategies for efficient intracellular uptake by adsorptive-mediated endocytosis, GST-fused proteins were cationized by forming a complex with a polycationic polyethylenimine (PEI)-glutathione conjugate. On screening of protein transduction, optimized PEI-glutathione conjugate for protein transduction was characterized by a partly oligomerized mixture of PEI with average molecular masses of 600 (PEI600) modified with multiple glutathiones, which could have sufficient avidity for GST. Furthermore, enhanced endosomal escape of transduced GST-fused proteins was observed when they were delivered with a glutathione-conjugated PEI600 derivative possessing a hydroxybutenyl moiety. These results were confirmed by both intracellular confocal imaging of GST-fused green fluorescent protein and activation of an endogenous growth signal transduction pathway by a GST-fused constitutively active mutant of a kinase protein. These PEI-glutathione conjugates seem to be convenient molecular tools for protein transduction of widely used GST-fused proteins.
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
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pubmed:status |
MEDLINE
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pubmed:month |
Oct
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pubmed:issn |
0021-924X
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pubmed:author |
pubmed-author:FutamiJunichiroJ,
pubmed-author:HuhNam-hoNH,
pubmed-author:KitazoeMidoriM,
pubmed-author:KosakaMegumiM,
pubmed-author:MurataHitoshiH,
pubmed-author:NakanishiHidetakaH,
pubmed-author:SakaguchiMasakiyoM,
pubmed-author:SenoMasaharuM,
pubmed-author:TadaHirokoH,
pubmed-author:YagiYasuyukiY,
pubmed-author:YamadaHidenoriH,
pubmed-author:YoneharaTakayukiT
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pubmed:issnType |
Print
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pubmed:volume |
144
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
447-55
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pubmed:dateRevised |
2009-11-19
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pubmed:meshHeading |
pubmed-meshheading:18603589-Animals,
pubmed-meshheading:18603589-BALB 3T3 Cells,
pubmed-meshheading:18603589-Biological Transport, Active,
pubmed-meshheading:18603589-CHO Cells,
pubmed-meshheading:18603589-COS Cells,
pubmed-meshheading:18603589-Cercopithecus aethiops,
pubmed-meshheading:18603589-Cricetinae,
pubmed-meshheading:18603589-Cricetulus,
pubmed-meshheading:18603589-Drug Carriers,
pubmed-meshheading:18603589-Endocytosis,
pubmed-meshheading:18603589-Glutathione,
pubmed-meshheading:18603589-Glutathione Transferase,
pubmed-meshheading:18603589-Green Fluorescent Proteins,
pubmed-meshheading:18603589-HeLa Cells,
pubmed-meshheading:18603589-Humans,
pubmed-meshheading:18603589-MAP Kinase Kinase 1,
pubmed-meshheading:18603589-Mice,
pubmed-meshheading:18603589-Molecular Weight,
pubmed-meshheading:18603589-Polyethyleneimine,
pubmed-meshheading:18603589-Recombinant Fusion Proteins,
pubmed-meshheading:18603589-Transduction, Genetic
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pubmed:year |
2008
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pubmed:articleTitle |
Intracellular delivery of glutathione S-transferase-fused proteins into mammalian cells by polyethylenimine-glutathione conjugates.
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pubmed:affiliation |
Department of Bioscience and Biotechnology, Faculty of Engineering, Graduate School of Natural Science and Technology, Okayama University, Okayama, Japan.
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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