Design of a protein tag and fluorogenic probe with modular structure for live-cell imaging of intracellular proteins

Yuko Kamikawa, Yuichiro Hori, Kazuo Yamashita, Lin Jin, Shinya Hirayama, Daron M. Standley, Kazuya Kikuchi

Research output: Contribution to journalArticlepeer-review

18 Citations (Scopus)

Abstract

Conditional fluorescence imaging is a powerful technique for precise spatiotemporal analysis of proteins in live cells upon administration of a synthetic probe. To be applicable to various biological phenomena, probes must be membrane-permeable, have a high signal-to-noise ratio, and work quickly. To date, few probes meet all of these requirements. Here, we designed a fluorogenic probe (AcFCANB) that could label intracellular proteins fused to the photoactive yellow protein (PYP) tag in live cells within 30 min and used it to image heterochromatin protein 1 localization in nuclei. AcFCANB is based on a modular platform consisting of fluorophore, ligand and quencher. We accelerated the labeling reaction by strategic mutations of charged residues on the surface of PYP. A simple model based on molecular dynamics simulations quantitatively reproduced the cooperative effect of multiple mutations on labeling rate.

Original languageEnglish
Pages (from-to)308-314
Number of pages7
JournalChemical Science
Volume7
Issue number1
DOIs
Publication statusPublished - Jan 1 2016
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Chemistry(all)

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