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Localization of Seed Oil Body Proteins in Tobacco Protoplasts Reveals Specific Mechanisms of Protein Targeting to Leaf Lipid Droplets

查看全文 作　　者：Stefania De [1]Domenico;Stefania [1]Bonsegna;Marcello Salvatore [2]Lenucci;Palmiro [1]Poltronieri;Gian Pietro Di [2]Sansebastiano;Angelo [1]Santino 高影响力作者 机构地区：[1]Institute of Sciences of Food Production C.N.R. Unit of Lecce, via Monteroni, Lecce 73100, Italy;[2]Dipartimento di Scienze e Tecnologie Biologiche ed Ambientali, Universita` del Salento, via Monteroni, Lecce 73100, Italy高影响力机构 出　　处：《Journal of Integrative Plant Biology》索引2011年第53卷第11期,共11页高影响力期刊 摘　　要：Oleosin, caleosin and steroleosin are normally expressed in developing seed cells and are targeted to oil bodies. In the present work, the cDNA of each gene tagged with fluorescent proteins was transiently expressed into tobacco protoplasts and the fluorescent patterns observed by confocal laser scanning microscopy. Our results indicated clear differences in the endocellular localization of the three proteins. Oleosin and caleosin both share a common structure consisting of a central hydrophobic domain flanked by two hydrophilic domains and were correctly targeted to lipid droplets (LD), whereas steroleosin, characterized by an N-terminal oil body anchoring domain, was mainly retained in the endoplasmic reticulum (ER). Protoplast fractionation on sucrose gradients indicated that both oleosin and caleosingreen fluorescent protein (GFP) peaked at different fractions than where steroleosin-GFP or the ER marker binding immunoglobulin protein (BiP), were recovered. Chemical analysis confirmed the presence of triacylglycerols in one of the fractions where oleosin-GFP was recovered. Finally, only oleosin- and caleosin-GFP were able to reconstitute artificial oil bodies in the presence of triacylglycerols and phospholipids. Taken together, our results pointed out for the first time that leaf LDs can be separated by the ER and both oleosin or caleosin are selectively targeted due to the existence of selective mechanisms controlling protein association with these organelles. 关 键 词：烟草原生质体 蛋白质 本地化 机制 机构 绿色荧光蛋白 共聚焦激光扫描 籽油
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