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

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 So...

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Hauptverfasser: Best, Marcel (VerfasserIn) , Porth, Isabel (VerfasserIn) , Hauke, Sebastian (VerfasserIn) , Braun, Felix (VerfasserIn) , Herten, Dirk-Peter (VerfasserIn) , Wombacher, Richard (VerfasserIn)
Dokumenttyp: Article (Journal)
Sprache:Englisch
Veröffentlicht: 07 Apr 2016
In: Organic & biomolecular chemistry
Year: 2016, Jahrgang: 14, Heft: 24, Pages: 5606-5611
ISSN:1477-0539
DOI:10.1039/C6OB00365F
Online-Zugang:Verlag, lizenzpflichtig, Volltext: https://doi.org/10.1039/C6OB00365F
Verlag, lizenzpflichtig, Volltext: https://pubs.rsc.org/en/content/articlelanding/2016/ob/c6ob00365f
Volltext
Verfasserangaben:Marcel Best, Isabel Porth, Sebastian Hauke, Felix Braun, Dirk-Peter Herten, Richard Wombacher
Beschreibung
Zusammenfassung: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.
Beschreibung:Gesehen am 11.05.2020
Beschreibung:Online Resource
ISSN:1477-0539
DOI:10.1039/C6OB00365F

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