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mRNA Cap Modification through Carbamate Chemistry: Synthesis of Amino‐ and Carboxy‐Functionalised Cap Analogues Suitable for Labelling and Bioconjugation
Author(s) -
Warminski Marcin,
Warminska Zofia,
Kowalska Joanna,
Jemielity Jacek
Publication year - 2015
Publication title -
european journal of organic chemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.825
H-Index - 155
eISSN - 1099-0690
pISSN - 1434-193X
DOI - 10.1002/ejoc.201500672
Subject(s) - chemistry , bioconjugation , moiety , stereochemistry , ribose , polymerase , amino acid , chemical modification , combinatorial chemistry , biochemistry , enzyme
A simple and efficient synthesis of dinucleotide mRNA 5′ cap analogues functionalised by the addition of an amine or carboxylic acid group onto one of the ribose rings is reported. Selection of the modification sites was dictated by the recognition specificity for the 5′ cap by various cap‐binding proteins, including eIF4E, DcpS, and Dcp2, as well as by bacteriophage RNA polymerase, which enables the incorporation of the synthetic cap into the mRNA chain. Some analogues were further modified by O‐to‐CH 2 substitution in the triphosphate bridge to confer resistance to specific hydrolases. Spatial crowding and structural rigidity were balanced by use of diamines and amino acids of different lengths attached to the ribose through a carbamate moiety. The impact of these substitutions on the conformational equilibria of the ribose components was investigated by 1 H NMR spectroscopy. The utility of the analogues for labelling with fluorescent dyes and affinity tags was demonstrated with the aid of NHS activation chemistry.