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

• Verfasst von: Best, Marcel [VerfasserIn]
• Verfasst von: Porth, Isabel [VerfasserIn]
• Verfasst von: Hauke, Sebastian [VerfasserIn]
• Verfasst von: Braun, Felix [VerfasserIn]
• Verfasst von: Herten, Dirk-Peter [VerfasserIn]
• Verfasst von: Wombacher, Richard [VerfasserIn]
• Titel: Protein-specific localization of a rhodamine-based calcium-sensor in living cells
• Verf.angabe: Marcel Best, Isabel Porth, Sebastian Hauke, Felix Braun, Dirk-Peter Herten, Richard Wombacher
• E-Jahr: 2016
• Jahr: 07 Apr 2016
• Umfang: 6 S.
• Fussnoten: Gesehen am 11.05.2020
• Titel Quelle: Enthalten in: Organic & biomolecular chemistry
• Ort Quelle: Cambridge : Royal Society of Chemistry, 2003
• Jahr Quelle: 2016
• Band/Heft Quelle: 14(2016), 24, Seite 5606-5611
• ISSN Quelle: 1477-0539
• DOI: doi:10.1039/C6OB00365F
• Datenträger: Online-Ressource
• Sprache: eng
• K10plus-PPN: 1697803075

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.