气候变化领域集成服务门户(CCPortal): Nanopore-mediated protein delivery enabling three-color single-molecule tracking in living cells

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DOI	10.1073/pnas.2012229118
	Nanopore-mediated protein delivery enabling three-color single-molecule tracking in living cells
	Chen Z.; Cao Y.; Lin C.-W.; Alvarez S.; Oh D.; Yang P.; Groves J.T.
发表日期	2021
ISSN	00278424
卷号	118 期号:5
英文摘要	Multicolor single-molecule tracking (SMT) provides a powerful tool to mechanistically probe molecular interactions in living cells. However, because of the limitations in the optical and chemical properties of currently available fluorophores and the multiprotein labeling strategies, intracellular multicolor SMT remains challenging for general research studies. Here, we introduce a practical method employing a nanopore-electroporation (NanoEP) technique to deliver multiple organic dye-labeled proteins into living cells for imaging. It can be easily expanded to three channels in commercial microscopes or be combined with other in situ labeling methods. Utilizing NanoEP, we demonstrate three-color SMT for both cytosolic and membrane proteins. Specifically, we simultaneously monitored single-molecule events downstream of EGFR signaling pathways in living cells. The results provide detailed resolution of the spatial localization and dynamics of Grb2 and SOS recruitment to activated EGFR along with the resultant Ras activation. © 2021 National Academy of Sciences. All rights reserved.
英文关键词	EGFR signaling; Electroporation; Ex vivo protein delivery; Nanopore; Single molecule
语种	英语
来源期刊	Proceedings of the National Academy of Sciences of the United States of America
文献类型	期刊论文
条目标识符	http://gcip.llas.ac.cn/handle/2XKMVOVA/180831
作者单位	Multiscale Research Institute of Complex Systems, Fudan University, Shanghai, 200433, China; Department of Chemistry, University of California, Berkeley, CA 94720, United States; Department of Materials Science and Engineering, University of California, Berkeley, CA 94720, United States; Mechanobiology Institute, National University of Singapore117411, Singapore
推荐引用方式 GB/T 7714	Chen Z.,Cao Y.,Lin C.-W.,et al. Nanopore-mediated protein delivery enabling three-color single-molecule tracking in living cells[J],2021,118(5).
APA	Chen Z..,Cao Y..,Lin C.-W..,Alvarez S..,Oh D..,...&Groves J.T..(2021).Nanopore-mediated protein delivery enabling three-color single-molecule tracking in living cells.Proceedings of the National Academy of Sciences of the United States of America,118(5).
MLA	Chen Z.,et al."Nanopore-mediated protein delivery enabling three-color single-molecule tracking in living cells".Proceedings of the National Academy of Sciences of the United States of America 118.5(2021).