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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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Publication details

The article was received on 30 Jun 2015, accepted on 28 Sep 2015 and first published on 30 Sep 2015


Article type: Edge Article
DOI: 10.1039/C5SC02351C
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Citation: Chem. Sci., 2016,7, 308-314
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    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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