Development of fluorogenic probes for quick No-Wash live-cell imaging of intracellular proteins

Yuichiro Hori, Tomoya Norinobu, Motoki Sato, Kyohei Arita, Masahiro Shirakawa, Kazuya Kikuchi

Research output: Contribution to journalArticlepeer-review

87 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)12360-12365
Number of pages6
JournalJournal of the American Chemical Society
Volume135
Issue number33
DOIs
Publication statusPublished - Aug 21 2013
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Catalysis
  • Chemistry(all)
  • Biochemistry
  • Colloid and Surface Chemistry

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