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| A modified GFP facilitates counting membrane protein subunits by step-wise photobleaching in Arabidopsis | |
Song, Kai1; Xue, Yiqun1; Wang, Xiaohua; Wan, Yinglang2; Deng, Xin ; Lin, Jinxing
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| 2017 | |
| 发表期刊 | JOURNAL OF PLANT PHYSIOLOGY
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| ISSN | 0176-1617 |
| 卷号 | 213页码:129-133 |
| 摘要 | Membrane proteins exert functions by forming oligomers or molecular complexes. Currently, step-wise photobleaching has been applied to count the fluorescently labelled subunits in plant cells, for which an accurate and reliable control is required to distinguish individual subunits and define the basal fluorescence. However, the common procedure using immobilized GFP molecules is obviously not applicable for analysis in living plant cells. Using the spatial intensity distribution analysis (SpIDA), we found that the A206K mutation reduced the dimerization of GFP molecules. Further ectopic expression of Myristoyl-GFP(A206K) driven by the endogenous AtCLC2 promoter allowed imaging of individual molecules at a low expression level. As a result, the percentage of dimers in the transgenic pCLC2::Myristoyl-mGFP(A206K) line was significantly reduced in comparison to that of the pCLC2::Myristoyl-GFP line, confirming its application in defining the basal fluorescence intensity of GFP. Taken together, our results demonstrated that pCLC2::Myristoyl-mGFP(A206K) can be used as a standard control for monomer GFP, facilitating the analysis of the step-wise photobleaching of membrane proteins in Arabidopsis thaliana. |
| 关键词 | Monomeric GFP VA-TIRFM Step-wise photobleaching Subunit counting Arabidopsis |
| 学科领域 | Plant Sciences |
| DOI | 10.3389/fpls.2017.00341 |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS关键词 | SINGLE-MOLECULE ; OLIGOMERIZATION ; DYNAMICS |
| WOS记录号 | WOS:000396080600001 |
| 出版者 | ELSEVIER GMBH |
| 文献子类 | Article |
| 出版地 | MUNICH |
| EISSN | 1618-1328 |
| 资助机构 | Major Science Foundation of the Ministry of Education of China (111 project) [313008] ; Programof Introducing Talents of Discipline to Universities (111 project)Ministry of Education, China - 111 Project [B13007] ; National Nature Science Foundation of ChinaNational Natural Science Foundation of China (NSFC) [31270224, 31270412] |
| 作者邮箱 | linjx@ibcas.ac.cn |
| 作品OA属性 | gold, Green Published |
| 引用统计 | |
| 文献类型 | 期刊论文 |
| 条目标识符 | http://ir.ibcas.ac.cn/handle/2S10CLM1/22008 |
| 专题 | 中科院北方资源植物重点实验室 |
| 作者单位 | 1.Chinese Acad Sci, Inst Bot, Key Lab Plant Resources, Beijing 100093, Peoples R China 2.Univ Chinese Acad Sci, Beijing 100049, Peoples R China 3.Beijing Forestry Univ, Coll Biol Sci & Biotechnol, Beijing 100083, Peoples R China |
| 推荐引用方式 GB/T 7714 | Song, Kai,Xue, Yiqun,Wang, Xiaohua,et al. A modified GFP facilitates counting membrane protein subunits by step-wise photobleaching in Arabidopsis[J]. JOURNAL OF PLANT PHYSIOLOGY,2017,213:129-133. |
| APA | Song, Kai,Xue, Yiqun,Wang, Xiaohua,Wan, Yinglang,Deng, Xin,&Lin, Jinxing.(2017).A modified GFP facilitates counting membrane protein subunits by step-wise photobleaching in Arabidopsis.JOURNAL OF PLANT PHYSIOLOGY,213,129-133. |
| MLA | Song, Kai,et al."A modified GFP facilitates counting membrane protein subunits by step-wise photobleaching in Arabidopsis".JOURNAL OF PLANT PHYSIOLOGY 213(2017):129-133. |
| 条目包含的文件 | ||||||
| 文件名称/大小 | 文献类型 | 版本类型 | 开放类型 | 使用许可 | ||
| fpls-08-00341.pdf（934KB） | 期刊论文 | 出版稿 | 开放获取 | CC BY-NC-SA | 浏览 请求全文 | |
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