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Issue 1, 2016
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Design of a protein tag and fluorogenic probe with modular structure for live-cell imaging of intracellular proteins

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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.

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

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Supplementary files

Article information


Submitted
30 Jun 2015
Accepted
28 Sep 2015
First published
30 Sep 2015

This article is Open Access
All publication charges for this article have been paid for by the Royal Society of Chemistry

Chem. Sci., 2016,7, 308-314
Article type
Edge Article
Author version available

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

Y. Kamikawa, Y. Hori, K. Yamashita, L. Jin, S. Hirayama, D. M. Standley and K. Kikuchi, Chem. Sci., 2016, 7, 308
DOI: 10.1039/C5SC02351C

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