Site-specific conjugation of 8-ethynyl-BODIPY to a protein by [2+3] cycloaddition

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dc.contributor.authorAlbrecht, Marcel
dc.contributor.authorLippach, Andreas
dc.contributor.authorExner, Matthias P.
dc.contributor.authorJerbi, Jihene
dc.contributor.authorSpringborg, Michael
dc.contributor.authorBudisa, Nediljko
dc.contributor.authorWenz, Gerhard
dc.date.accessioned2017-10-25T06:27:20Z
dc.date.available2017-10-25T06:27:20Z
dc.date.issued2015
dc.description.abstractWe 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.en
dc.identifier.eissn1477-0539
dc.identifier.issn1477-0520
dc.identifier.pmid25994282
dc.identifier.urihttps://depositonce.tu-berlin.de/handle/11303/6953
dc.identifier.urihttp://dx.doi.org/10.14279/depositonce-6292
dc.language.isoen
dc.rights.urihttps://creativecommons.org/licenses/by/3.0/
dc.subject.ddc540 Chemie und zugeordnete Wissenschaftende
dc.titleSite-specific conjugation of 8-ethynyl-BODIPY to a protein by [2+3] cycloadditionen
dc.typeArticleen
dc.type.versionpublishedVersionen
dcterms.bibliographicCitation.doi10.1039/c5ob00505a
dcterms.bibliographicCitation.issue24
dcterms.bibliographicCitation.journaltitleOrganic & biomolecular chemistry : OBCen
dcterms.bibliographicCitation.originalpublishernameRoyal Society of Chemistryde
dcterms.bibliographicCitation.originalpublisherplaceCambridgede
dcterms.bibliographicCitation.pageend6736
dcterms.bibliographicCitation.pagestart6728
dcterms.bibliographicCitation.volume13
tub.accessrights.dnbfree
tub.affiliationFak. 2 Mathematik und Naturwissenschaften::Inst. Chemie::FG Biokatalysede
tub.affiliation.facultyFak. 2 Mathematik und Naturwissenschaftende
tub.affiliation.groupFG Biokatalysede
tub.affiliation.instituteInst. Chemiede
tub.publisher.universityorinstitutionTechnische Universität Berlin

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