Development of Fluorogenic Probes for Quick No-Wash Live-Cell Imaging of Intracellular Proteins

作者：Hori Yuichiro; Norinobu Tomoya; Sato Motoki; Arita Kyohei; Shirakawa Masahiro; Kikuchi Kazuya^*

来源：Journal of the American Chemical Society, 2013, 135(33): 12360-12365.

DOI：10.1021/ja405745v

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.

• 出版日期2013-8-21