A Bifunctional Fluorogenic Rhodamine Probe for Proximity‐Induced Bioorthogonal Chemistry. Issue 72 (28th November 2017)

• Record Type: Journal Article
• Authors: Werther, Philipp; Möhler, Jasper S.; Wombacher, Richard
• Is Part Of: Chemistry. Volume 23:Issue 72(2017)
• Page Start: 18216
• Page End: 18224
• Publication Date: 2017-11-28
• Subjects: dienophiles -- kinetics -- proteins -- rhodamine -- tetrazine; Chemistry -- Periodicals; 540
• DOI: 10.1002/chem.201703607
• Languages: English
• ISSNs: 0947-6539

Abstract

Abstract: Bioorthogonal reactions have emerged as a versatile tool in life sciences. The inverse electron demand Diels–Alder reaction (DAinv ) stands out due to the availability of reactants with very fast kinetics. However, highly reactive dienophiles suffer the disadvantage of being less stable and prone to side reactions. Herein, we evaluate the extent of acceleration of rather unreactive but highly stable dienophiles by DNA‐templated proximity. To this end, we developed a modular synthetic route for a novel bifunctional fluorogenic tetrazine rhodamine probe that we used to determine the reaction kinetics of various dienophiles in a fluorescence assay. Under proximity‐driven conditions the reaction was found to be several orders of magnitude faster, and we observed almost no background reaction when proximity was not induced. This fundamental study identifies a minimally sized fluorogenic tetrazine dienophile reactant pair that has potential to be generally used for the visualization of biomolecular interactions with temporal and spatial resolution in living systems. Abstract : Let there be light ! A novel bifunctional rhodamine reporter is presented. Conversion of the attached tetrazine group in Diels‐Alder reactions results in "light‐up" of fluorescence. This property was used to study the effect of proximity on the reactivity of various dienophiles in Diels‐Alder reactions. Moreover, the bifunctional dye enables to transfer biomolecular interactions to a fluorescence signal.