In Vitro Protein Import of a Putative Amino Acid Transporter from Arabidopsis thaliana into Chloroplasts and Its Suborganellar Localization
In Vitro Protein Import of a Putative Amino Acid Transporter from Arabidopsis thaliana into Chloroplasts and Its Suborganellar Localization
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DOI：
10.1271/bbb.110489
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发表时间：
2011-11-01
影响因子：
1.6
通讯作者：
Akita, Mitsuru
中科院分区：
文献类型：
--
作者：
Sattasuk, Kwanchanok;Nozawa, Akira;Akita, Mitsuru
我们从拟南芥中鉴定了At 5g 19500（At 5g 19500 p）的基因产物，其与大肠杆菌的酪氨酸特异性转运蛋白EcTyrP同源。对At 5g 19500 p的氨基酸序列的计算分析预测了11个跨膜结构域（TMD）和在其氨基末端的潜在质体靶向信号。作为理解At 5g 19500 p在植物细胞中可能作用的第一步，我们尝试通过体外叶绿体输入测定法使用在无细胞小麦胚芽系统中翻译的At 5g 19500 p来确定At 5g 19500 p的定位（Madin埃塔尔，Proc. Natl. Acad. Sci. USA，97，559 -564（2000）），随后进行叶绿体的亚分级分离。At 5g 19500 p被成功导入叶绿体，并从内被膜回收新转运的成熟形式At 5g 19500 p。
We identified a gene product of At5g19500 (At5g19500p) from Arabidopsis thaliana that is homologous to EcTyrP, a tyrosine-specific transporter from Escherichia coli. Computational analyses of the amino acid sequence of At5g19500p predicted 11 transmembrane domains (TMDs) and a potential plastid targeting signal at its amino terminus. As a first step toward understanding the possible role of At5g19500p in plant cells, we attempted to determine the localization of At5g19500p by an in vitro chloroplastic import assay using At5g19500p translated in a cell-free wheat germ system (Madin etal., Proc. Natl. Acad. Sci. USA, 97,559-564 (2000)), followed by subfractionation of the chloroplasts. At5g19500p was successfully imported into chloroplasts, and the newly transported mature form of At5g19500p was recovered from the inner envelope membrane.