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Title: Live-cell imaging of membrane proteins by a coiled-coil labeling method—Principles and applications
Authors: Yano, Yoshiaki
Matsuzaki, Katsumi
Author's alias: 矢野, 義明
松﨑, 勝巳
Keywords: Coiled-coil
Fluorescence imaging
Membrane protein
Tag–probe labeling
Issue Date: 1-May-2019
Publisher: Elsevier B.V.
Journal title: Biochimica et Biophysica Acta - Biomembranes
Volume: 1861
Issue: 5
Start page: 1011
End page: 1017
Abstract: In situ investigations in living cell membranes are important to elucidate the dynamic behaviors of membrane proteins in complex biomembrane environments. Protein-specific labeling is a key technique for the detection of a target protein by fluorescence imaging. The use of post-translational labeling methods using a genetically encodable tag and synthetic probes targeting the tag offer a smaller label size, labeling with synthetic fluorophores, and precise control of the labeling ratio in multicolor labeling compared with conventional genetic fusions with fluorescent proteins. This review focuses on tag–probe labeling studies for live-cell analysis of membrane proteins based on heterodimeric peptide pairs that form coiled-coil structures. The robust and simple peptide–peptide interaction enables not only labeling of membrane proteins by noncovalent interactions, but also covalent crosslinking and acyl transfer reactions guided by coiled-coil assembly. A number of studies have demonstrated that membrane protein behaviors in live cells, such as internalization of receptors and the oligomeric states of various membrane proteins (G-protein-coupled receptors, epidermal growth factor receptors, influenza A M2 channel, and glycopholin A), can be precisely analyzed using coiled-coil labeling, indicating the potential of this labeling method in membrane protein research.
Rights: © 2019. This manuscript version is made available under the CC-BY-NC-ND 4.0 license .
The full-text file will be made open to the public on 1 May 2020 in accordance with publisher's 'Terms and Conditions for Self-Archiving'.
This is not the published version. Please cite only the published version. この論文は出版社版でありません。引用の際には出版社版をご確認ご利用ください。
DOI(Published Version): 10.1016/j.bbamem.2019.02.009
PubMed ID: 30831076
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