Department of Molecular Biotechnology and Kumho Life Science Laboratory, College of Agriculture and Life Sciences, Chonnam National University, Gwangju 500-757, Republic of Korea.
Plant Sci. 2011 Sep;181(3):242-8. doi: 10.1016/j.plantsci.2011.05.016. Epub 2011 Jun 14.
The dual positional maize lipoxygenase-1 was introduced into rice and T2 transgenic plants were produced. Cellular location of maize lipoxygenase-1 in transgenic rice and effects of calcium ion on membrane association in vitro were analyzed. Localization study by confocal microscopic analysis indicated that the maize lipoxygenase-1 was localized in cytoplasm. Sucrose-density fractionation experiment and in vitro protein transport to chloroplast showed that the maize lipoxygenase-1 can be associated with chloroplast. Secondary structure alignment revealed putative calcium binding sites in the PLAT domain of maize lipoxygenase-1 and the association of the maize lipoxygenase-1 with membranes was mediated by calcium ion in vitro. Our results provide evidences for calcium-mediated translocation of dual positional LOX without chloroplast targeting sequence from cytoplasm to chloroplast in plants for the first time.
将双定位玉米脂氧合酶-1导入水稻中,并产生了 T2 转基因植物。分析了转基因水稻中玉米脂氧合酶-1的细胞定位以及钙离子对体外膜结合的影响。通过共焦显微镜分析的定位研究表明,玉米脂氧合酶-1定位于细胞质中。蔗糖密度分级实验和体外蛋白向叶绿体的转运表明,玉米脂氧合酶-1可以与叶绿体结合。二级结构比对显示,玉米脂氧合酶-1的 PLAT 结构域中存在潜在的钙结合位点,体外实验表明,钙离子介导了玉米脂氧合酶-1与膜的结合。我们的研究结果首次为植物中没有叶绿体靶向序列的双定位 LOX 从细胞质向叶绿体的钙介导易位提供了证据。