Single-molecule fluorescence imaging to quantify membrane protein dynamics and oligomerization in living plant cells

doi:10.1038/nprot.2015.132

	Single-molecule fluorescence imaging to quantify membrane protein dynamics and oligomerization in living plant cells
	Wang, Xiaohua ; Li, Xiaojuan 1; Deng, Xin; Luu, Doan-Trung ; Maurel, Christophe 2; Lin, Jinxing 1
	2015
发表期刊	NATURE PROTOCOLS
ISSN	1754-2189
卷号	10 期号:12 页码:2054-2063
摘要	Measuring the mobility and interactions of proteins is key to understanding cellular signaling mechanisms; however, quantitative analysis of protein dynamics in living plant cells remains a major challenge. Here we describe an automated, single-molecule protocol based on total internal reflection fluorescence microscopy (TIRFM) imaging that allows protein tracking and subunit counting in living plant cells. This protocol uses TIRFM to image transgenic plant tissues expressing fluorescently tagged proteins that are localized to the plasma membrane. Next, a tracking algorithm quantifies dynamic changes in fluorescent protein motion types, temporary particle displacement and protein photobleaching steps. This protocol allows researchers to study the kinetic characteristics of heterogeneously distributed proteins. The approach has potential applications for studies of protein dynamics and subunit stoichiometry for a wide variety of plasma membrane and intracellular proteins in living plant cells and other biological specimens visualized by TIRFM or other fluorescence imaging techniques. The whole protocol can be completed in 5-6 h.
学科领域	Biochemical Research Methods
DOI	10.1038/nprot.2015.132
收录类别	SCI
语种	英语
WOS关键词	PARTICLE TRACKING ; PLASMA-MEMBRANE ; ARABIDOPSIS ; CLATHRIN ; MICRODOMAINS ; MICROSCOPY ; DIFFUSION ; SEQUENCES ; REVEALS
WOS研究方向	Science Citation Index Expanded (SCI-EXPANDED)
WOS记录号	WOS:000366122600011
出版者	NATURE PUBLISHING GROUP
文献子类	Article
出版地	LONDON
EISSN	1750-2799
资助机构	Program of Introducing Talents of Discipline to Universities (111 project) [B13007] ; Major Science Foundation of the Ministry of Education of China [313008] ; National Basic Research Program of China (973 Program) [2011CB809103] ; National Nature Science Foundation of China Project [31270412, 31270224]
作者邮箱	linjinxing@bjfu.edu.cn
引用统计	被引频次：55[WOS] [WOS记录] [WOS相关记录]
文献类型	期刊论文
条目标识符	http://ir.ibcas.ac.cn/handle/2S10CLM1/25691
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, Peoples R China 2.Beijing Forestry Univ, Coll Biol Sci & Technol, Minist Educ, Key Lab Genet & Breeding Forest Trees & Ornament, Beijing, Peoples R China 3.Univ Montpellier 2, Inst Natl Rech Agron Montpellier SupAgro, Inst Biol Integrat Plantes, CNRS,UMR 5004,UMR 0386,Biochim & Physiol Mol Plan, Montpellier, France
推荐引用方式 GB/T 7714	Wang, Xiaohua,Li, Xiaojuan,Deng, Xin,et al. Single-molecule fluorescence imaging to quantify membrane protein dynamics and oligomerization in living plant cells[J]. NATURE PROTOCOLS,2015,10(12):2054-2063.
APA	Wang, Xiaohua,Li, Xiaojuan,Deng, Xin,Luu, Doan-Trung,Maurel, Christophe,&Lin, Jinxing.(2015).Single-molecule fluorescence imaging to quantify membrane protein dynamics and oligomerization in living plant cells.NATURE PROTOCOLS,10(12),2054-2063.
MLA	Wang, Xiaohua,et al."Single-molecule fluorescence imaging to quantify membrane protein dynamics and oligomerization in living plant cells".NATURE PROTOCOLS 10.12(2015):2054-2063.

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