Installation of authentic BicA and SbtA proteins to the chloroplast envelope membrane is achieved by the proteolytic cleavage of chimeric proteins in Arabidopsis
Installation of authentic BicA and SbtA proteins to the chloroplast envelope membrane is achieved by the proteolytic cleavage of chimeric proteins in Arabidopsis
复制标题
通过拟南芥中嵌合蛋白的蛋白水解切割，将真正的 BicA 和 SbtA 蛋白安装到叶绿体包膜上
DOI：
10.1038/s41598-020-59190-1
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发表时间：
2020
影响因子：
4.6
通讯作者：
Inaba Takehito
中科院分区：
文献类型：
--
作者：
Uehara Susumu;Sei Ayane;Sada Misaki;Ito-Inaba Yasuko;Inaba Takehito
为了提高C3植物的光合性能，多年来一直有人提出在叶绿体内被膜上安装蓝藻碳酸氢盐转运蛋白。在我们之前的研究中，我们成功地将嵌合蓝藻钠依赖性碳酸氢盐转运蛋白BicA或SbtA引入拟南芥叶绿体IEM。然而，真正的BicA和SbtA的叶绿体IEM的安装尚未实现。在这项研究中，我们研究了烟草蚀纹病毒（TEV）蛋白酶是否靶向叶绿体内可以切割嵌合蛋白，并产生真正的碳酸氢盐转运蛋白。为此，我们构建了携带转运肽的TEV蛋白酶，并将其与含有TEV切割位点的嵌合BicA或SbtA蛋白在植物中表达。嵌合蛋白仅在TEV蛋白酶共表达时才被切割。在叶绿体IEM中检测到血凝素标记的BicA和SbtA的真实形式。此外，切割嵌合蛋白在TEV识别位点似乎发生后，嵌合蛋白的叶绿体IEM的目标。我们得出结论，叶绿体内嵌合蛋白的切割是一种有效的方式来安装真正的碳酸氢盐转运蛋白的叶绿体IEM。此外，类似的方法可以应用于其他细菌质膜蛋白。
To improve the photosynthetic performance of C3plants, installing cyanobacterial bicarbonate transporters to the chloroplast inner envelope membrane (IEM) has been proposed for years. In our previous study, we successfully introduced chimeric cyanobacterial sodium-dependent bicarbonate transporters, BicA or SbtA, to the chloroplast IEM of Arabidopsis. However, the installation of authentic BicA and SbtA to the chloroplast IEM has not been achieved yet. In this study, we examined whether or not tobacco etch virus (TEV) protease targeted within chloroplasts can cleave chimeric proteins and produce authentic bicarbonate transporters. To this end, we constructed a TEV protease that carried the transit peptide and expressed it with chimeric BicA or SbtA proteins containing a TEV cleavage sitein planta. Chimeric proteins were cleaved only when the TEV protease was co-expressed. The authentic forms of hemagglutinin-tagged BicA and SbtA were detected in the chloroplast IEM. In addition, cleavage of chimeric proteins at the TEV recognition site seemed to occur after the targeting of chimeric proteins to the chloroplast IEM. We conclude that the cleavage of chimeric proteins within chloroplasts is an efficient way to install authentic bicarbonate transporters to the chloroplast IEM. Furthermore, a similar approach can be applied to other bacterial plasma membrane proteins.