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Issue 24, 2016
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Protein-specific localization of a rhodamine-based calcium-sensor in living cells

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Abstract

A small synthetic calcium sensor that can be site-specifically coupled to proteins in living cells by utilizing the bio-orthogonal HaloTag labeling strategy is presented. We synthesized an iodo-derivatized BAPTA chelator with a tetramethyl rhodamine fluorophore that allows further modification by Sonogashira cross-coupling. The presented calcium sensitive dye shows a 200-fold increase in fluorescence upon calcium binding. The derivatization with an aliphatic linker bearing a terminal haloalkane-function by Sonogashira cross-coupling allows the localization of the calcium sensor to Halo fusion proteins which we successfully demonstrate in in vitro and in vivo experiments. The herein reported highly sensitive tetramethyl rhodamine based calcium indicator, which can be selectively localized to proteins, is a powerful tool to determine changes in calcium levels inside living cells with spatiotemporal resolution.

Graphical abstract: Protein-specific localization of a rhodamine-based calcium-sensor in living cells

  • This article is part of the themed collection: New Talent
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Supplementary files

Article information


Submitted
15 Feb 2016
Accepted
06 Apr 2016
First published
07 Apr 2016

This article is Open Access

Org. Biomol. Chem., 2016,14, 5606-5611
Article type
Communication

Protein-specific localization of a rhodamine-based calcium-sensor in living cells

M. Best, I. Porth, S. Hauke, F. Braun, D. Herten and R. Wombacher, Org. Biomol. Chem., 2016, 14, 5606
DOI: 10.1039/C6OB00365F

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