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Expanding the chemical scope of RNA: methyltransferases to site-specific alkynylation of RNA for click labeling

This work identifies the combination of enzymatic transfer and click labeling as an efficient method for the site-specific tagging of RNA molecules for biophysical studies. A double-activated analog of the ubiquitous co-substrate S -adenosyl- l -methionine was employed to enzymatically transfer a fi...

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Bibliographische Detailangaben
Hauptverfasser: Motorin, Yuri (VerfasserIn) , Burhenne, Jürgen (VerfasserIn) , Teimer, Roman (VerfasserIn) , Koynov, Kaloian (VerfasserIn) , Willnow, Sophie (VerfasserIn) , Weinhold, Elmar (VerfasserIn) , Helm, Mark (VerfasserIn)
Dokumenttyp: Article (Journal)
Sprache:Englisch
Veröffentlicht: 2011
In: Nucleic acids research
Year: 2011, Jahrgang: 39, Heft: 5, Pages: 1943-1952
ISSN:1362-4962
DOI:10.1093/nar/gkq825
Online-Zugang:Verlag, lizenzpflichtig, Volltext: https://doi.org/10.1093/nar/gkq825
Volltext
Verfasserangaben:Yuri Motorin, Jürgen Burhenne, Roman Teimer, Kaloian Koynov, Sophie Willnow, Elmar Weinhold and Mark Helm
Beschreibung
Zusammenfassung:This work identifies the combination of enzymatic transfer and click labeling as an efficient method for the site-specific tagging of RNA molecules for biophysical studies. A double-activated analog of the ubiquitous co-substrate S -adenosyl- l -methionine was employed to enzymatically transfer a five carbon chain containing a terminal alkynyl moiety onto RNA. The tRNA:methyltransferase Trm1 transferred the extended alkynyl moiety to its natural target, the N2 of guanosine 26 in tRNA Phe . LC/MS and LC/MS/MS techniques were used to detect and characterize the modified nucleoside as well as its cycloaddition product with a fluorescent azide. The latter resulted from a labeling reaction via Cu(I)-catalyzed azide-alkyne 1,3-cycloaddition click chemistry, producing site-specifically labeled RNA whose suitability for single molecule fluorescence experiments was verified in fluorescence correlation spectroscopy experiments.
Beschreibung:First published online: 30 October 2010
Gesehen am 08.09.2022
Beschreibung:Online Resource
ISSN:1362-4962
DOI:10.1093/nar/gkq825

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