Development of Fluorogenic Probes for Quick No-Wash Live-Cell Imaging of Intracellular Proteins
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- Yuichiro Hori
- JST, PRESTO, Suita, Osaka 565-0871, Japan
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- Kyohei Arita
- Graduate School of Medical Life Science, Yokohama City University, Yokohama, Kanagawa 230-0045, Japan
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- Masahiro Shirakawa
- Graduate School of Engineering, Kyoto University, Kyoto 615-8510, Japan
Description
We developed novel fluorogenic probes for no-wash live-cell imaging of proteins fused to PYP-tag, which is a small protein tag recently reported by our group. Through the design of a new PYP-tag ligand, specific intracellular protein labeling with rapid kinetics and fluorogenic response was accomplished. The probes crossed the cell membrane, and cytosolic and nuclear localizations of PYP-tagged proteins without cell washing were visualized within a 6-min reaction time. The fluorogenic response was due to the environmental effect of fluorophore upon binding to PYP-tag. Furthermore, the PYP-tag-based method was applied to the imaging of methyl-CpG-binding domain localization. This rapid protein-labeling system combined with the small protein tag and designed fluorogenic probes offers a powerful method to study the localization, movement, and function of cellular proteins.
Journal
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- Journal of the American Chemical Society
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Journal of the American Chemical Society 135 (33), 12360-12365, 2013-08-08
American Chemical Society (ACS)