Autoren:	Kellner, Stefanie Kollar, Laura Bettina Ochel, Antonia Ghate, Manjunath Helm, Mark
Titel:	Structure-function relationship of substituted bromomethylcoumarins in nucleoside specificity of RNA alkylation
Online-Publikationsdatum:	5-Okt-2022
Erscheinungsdatum:	2013
Sprache des Dokuments:	Englisch
Zusammenfassung/Abstract:	Selective alkylation of RNA nucleotides is an important field of RNA biochemistry, e.g. in applications of fluorescent labeling or in structural probing experiments, yet detailed structure-function studies of labeling agents are rare. Here, bromomethylcoumarins as reactive compounds for fluorescent labeling of RNA are developed as an attractive scaffold on which electronic properties can be modulated by varying the substituents. Six different 4-bromomethyl-coumarins of various substitution patterns were tested for nucleotide specificity of RNA alkylation using tRNA from Escherichia coli as substrate. Using semi-quantitative LC-MS/MS analysis, reactions at mildly acidic and slightly alkaline pH were compared. For all tested compounds, coumarin conjugates with 4-thiouridine, pseudouridine, guanosine, and uridine were identified, with the latter largely dominating. This data set shows that selectivity of ribonucleotide alkylation depends on the substitution pattern of the reactive dye, and even more strongly on the modulation of the reaction conditions. The latter should be therefore carefully optimized when striving to achieve selectivity. Interestingly, the highest selectivity for labeling of a modified nucleoside, namely of 4-thiouridine, was achieved with a compound whose selectivity was somewhat less dependent on reaction conditions than the other compounds. In summary, bromomethylcoumarin derivatives are a highly interesting class of compounds, since their selectivity for 4-thiouridine can be efficiently tuned by variation of substitution pattern and reaction conditions.
DDC-Sachgruppe:	500 Naturwissenschaften 500 Natural sciences and mathematics
Veröffentlichende Institution:	Johannes Gutenberg-Universität Mainz
Organisationseinheit:	FB 09 Chemie, Pharmazie u. Geowissensch.
Veröffentlichungsort:	Mainz
ROR:	https://ror.org/023b0x485
DOI:	http://doi.org/10.25358/openscience-7832
Version:	Published version
Publikationstyp:	Zeitschriftenaufsatz
Nutzungsrechte:	CC BY
Informationen zu den Nutzungsrechten:	https://creativecommons.org/licenses/by/3.0/
Zeitschrift:	PLoS one 8 7
Seitenzahl oder Artikelnummer:	e67945
Verlag:	PLoS
Verlagsort:	Lawrence, Kan.
Erscheinungsdatum:	2013
ISSN:	1932-6203
URL der Originalveröffentlichung:	http://dx.doi.org/10.1371/journal.pone.0067945
DOI der Originalveröffentlichung:	10.1371/journal.pone.0067945
Enthalten in den Sammlungen:	DFG-OA-Publizieren (2012 - 2017)