A modified GFP facilitates counting membrane protein subunits by step-wise photobleaching in Arabidopsis

doi:10.3389/fpls.2017.00341

CORC > 植物研究所 > 中国科学院植物研究所 > 中科院北方资源植物重点实验室

	A modified GFP facilitates counting membrane protein subunits by step-wise photobleaching in Arabidopsis
	Song, Kai 3; Xue, Yiqun 3; Wang, Xiaohua ; Wan, Yinglang 1; Deng, Xin ; Lin, Jinxing
刊名	JOURNAL OF PLANT PHYSIOLOGY
	2017
卷号	213 页码:129-133
关键词	Monomeric GFP VA-TIRFM Step-wise photobleaching Subunit counting Arabidopsis
ISSN号	0176-1617
DOI	10.3389/fpls.2017.00341
文献子类	Article
英文摘要	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.
学科主题	Plant Sciences
电子版国际标准刊号	1618-1328
出版地	MUNICH
WOS关键词	SINGLE-MOLECULE ; OLIGOMERIZATION ; DYNAMICS
语种	英语
出版者	ELSEVIER GMBH
WOS记录号	WOS:000396080600001
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]
内容类型	期刊论文
源URL	[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.Univ Chinese Acad Sci, Beijing 100049, Peoples R China 2.Beijing Forestry Univ, Coll Biol Sci & Biotechnol, Beijing 100083, Peoples R China 3.Chinese Acad Sci, Inst Bot, Key Lab Plant Resources, Beijing 100093, 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.