Single-molecule fluorescence imaging to quantify membrane protein dynamics and oligomerization in living plant cells
文献类型：期刊论文
| 作者 | Wang, Xiaohua; Li, Xiaojuan3; Deng, Xin ; Luu, Doan-Trung; Maurel, Christophe2; Lin, Jinxing3
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| 刊名 | NATURE PROTOCOLS
 |
| 出版日期 | 2015 |
| 卷号 | 10期号:12页码:2054-2063 |
| ISSN号 | 1754-2189 |
| DOI | 10.1038/nprot.2015.132 |
| 文献子类 | Article |
| 英文摘要 | 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. |
| 学科主题 | Biochemical Research Methods |
| 出版地 | LONDON |
| 电子版国际标准刊号 | 1750-2799 |
| 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) |
| 语种 | 英语 |
| WOS记录号 | WOS:000366122600011 |
| 出版者 | NATURE PUBLISHING GROUP |
| 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] |
| 源URL | [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.Univ Montpellier 2, Inst Natl Rech Agron Montpellier SupAgro, Inst Biol Integrat Plantes, CNRS,UMR 5004,UMR 0386,Biochim & Physiol Mol Plan, Montpellier, France 2.Beijing Forestry Univ, Coll Biol Sci & Technol, Minist Educ, Key Lab Genet & Breeding Forest Trees & Ornament, Beijing, Peoples R China 3.Chinese Acad Sci, Inst Bot, Key Lab Plant Resources, Beijing, Peoples R China |
| 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. |
入库方式： OAI收割
来源：植物研究所
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