Key Laboratory of Forest Tree Genetic Improvement and Biotechnology, Ministry of Education, Northeast Forestry University, 26 Hexing Road, Harbin 150040, China.
Mol Biol Rep. 2011 Jun;38(5):3045-54. doi: 10.1007/s11033-010-9971-y. Epub 2010 Feb 5.
The chloroplast is one of the most important organelles in plants. Proteomic investigations of chloroplasts have been undertaken for many herb plant species, but to date no such investigation has been reported for woody plant chloroplasts. In the present study we initiated a systematic proteomic study of Populus chloroplasts using a shotgun proteomic method. After isolation of chloroplasts and tryptic digestion of the proteins, the protein fragments were separated via HPLC using an SCX column, and the peptides were analyzed by LC-MS/MS; 119 proteins were successfully identified. Based on annotation information in the UniProtKB/Swiss-Prot database, these proteins were identified as being localized in the chloroplast thylakoid membrane, chloroplast stroma, chloroplast thylakoid lumen, and plastoglobules. Over 50% of all identified proteins were confirmed as chloroplast thylakoid proteins, and 85 are encoded by the chloroplast genome with the remaining proteins encoded by the nuclear genome. Based on functional annotation, these proteins were classified into four functional categories, including photosynthesis, redox regulation and stress, primary and secondary metabolism, transport and signaling. These data provide a valuable basis for further studies on photosynthesis in poplar species.
叶绿体是植物中最重要的细胞器之一。已经对许多草本植物物种的叶绿体进行了蛋白质组学研究，但迄今为止，尚未有木本植物叶绿体的此类研究报告。在本研究中，我们使用鸟枪法蛋白质组学方法对杨树叶绿体进行了系统的蛋白质组学研究。在叶绿体分离和蛋白质的胰酶消化后，使用 SCX 柱通过 HPLC 分离蛋白片段，并通过 LC-MS/MS 进行分析；成功鉴定了 119 种蛋白质。根据 UniProtKB/Swiss-Prot 数据库中的注释信息，这些蛋白质被鉴定为定位于叶绿体类囊体膜、叶绿体基质、叶绿体类囊体腔和质体小球中。鉴定出的所有蛋白质中有超过 50%被确认为叶绿体类囊体蛋白，其中 85 种由叶绿体基因组编码，其余蛋白质由核基因组编码。根据功能注释，这些蛋白质被分为四个功能类别，包括光合作用、氧化还原调节和应激、初级和次级代谢、运输和信号转导。这些数据为进一步研究杨树物种的光合作用提供了有价值的基础。
