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Main Title: Site-specific conjugation of 8-ethynyl-BODIPY to a protein by [2+3] cycloaddition
Author(s): Albrecht, Marcel
Lippach, Andreas
Exner, Matthias P.
Jerbi, Jihene
Springborg, Michael
Budisa, Nediljko
Wenz, Gerhard
Type: Article
Language Code: en
Abstract: We report a straightforward synthesis of 8-ethynyl-BODIPY derivatives and their potential as fluorescent labeling compounds using an alkyne-azide click chemistry approach. The ethynyl substituted BODIPY dyes at the meso-position were reacted under Cu+ catalysis and mild physiological conditions in organic and biological model systems using benzyl azide and a Barstar protein which was selectively modified by a single amino acid substituted methionine at the N-terminus (Met1). azidohomoalanine (Aha). Conjugation with the protein and the model azide was indicated by a significant blue shift upon formation of the triazole moiety system, which allowed easy distinction between free and coupled dyes. This blue shift was rationalized by the perpendicular orientation of the triazole relative to the chromophore using time dependent density functional theory (TDDFT) calculations. A full spectroscopic and thermodynamic characterization of the protein revealed that a fluorophore was incorporated without the cross influence of protein stability and functional integrity. Furthermore, model reactions of 8-ethynyl-BODIPY derivatives with benzyl azide under copper-free conditions indicate second order kinetics with high rate constants comparable with those found for the strain-promoted azide-alkyne cycloaddition (SPAAC). In this way, we establish a unique and highly efficient method to introduce alkyne-BODIPY into a protein scaffold potentially useful for diverse applications in areas ranging from fundamental protein dynamics studies to biotechnology or cell biology.
Issue Date: 2015
Date Available: 25-Oct-2017
DDC Class: 540 Chemie und zugeordnete Wissenschaften
Journal Title: Organic & biomolecular chemistry : OBC
Publisher: Royal Society of Chemistry
Publisher Place: Cambridge
Volume: 13
Issue: 24
Publisher DOI: 10.1039/c5ob00505a
Page Start: 6728
Page End: 6736
EISSN: 1477-0539
ISSN: 1477-0520
Appears in Collections:FG Biokatalyse » Publications

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