Unknown

Dataset Information

0

Quantum Chemical Calculations and Experimental Validation of the Photoclick Reaction for Fluorescent Labeling of the 5' cap of Eukaryotic mRNAs.


ABSTRACT: Bioorthogonal click reactions are powerful tools to specifically label biomolecules in living cells. Considerable progress has been made in site-specific labeling of proteins and glycans in complex biological systems, but equivalent methods for mRNAs are rare. We present a chemo-enzymatic approach to label the 5' cap of eukaryotic mRNAs using a bioorthogonal photoclick reaction. Herein, the N7-methylated guanosine of the 5' cap is enzymatically equipped with an allyl group using a variant of the trimethylguanosine synthase 2 from Giardia lamblia (GlaTgs2). To elucidate whether the resulting N (2)-modified 5' cap is a suitable dipolarophile for photoclick reactions, we used Kohn-Sham density functional theory (KS-DFT) and calculated the HOMO and LUMO energies of this molecule and nitrile imines. Our in silico studies suggested that combining enzymatic allylation of the cap with subsequent labeling in a photoclick reaction was feasible. This could be experimentally validated. Our approach generates a turn-on fluorophore site-specifically at the 5' cap and therefore presents an important step towards labeling of eukaryotic mRNAs in a bioorthogonal manner.

SUBMITTER: Stummer D 

PROVIDER: S-EPMC4522179 | biostudies-literature | 2015 Jun

REPOSITORIES: biostudies-literature

altmetric image

Publications

Quantum Chemical Calculations and Experimental Validation of the Photoclick Reaction for Fluorescent Labeling of the 5' cap of Eukaryotic mRNAs.

Stummer Daniela D   Herrmann Carmen C   Rentmeister Andrea A  

ChemistryOpen 20150204 3


Bioorthogonal click reactions are powerful tools to specifically label biomolecules in living cells. Considerable progress has been made in site-specific labeling of proteins and glycans in complex biological systems, but equivalent methods for mRNAs are rare. We present a chemo-enzymatic approach to label the 5' cap of eukaryotic mRNAs using a bioorthogonal photoclick reaction. Herein, the N7-methylated guanosine of the 5' cap is enzymatically equipped with an allyl group using a variant of the  ...[more]

Similar Datasets

| S-EPMC9376930 | biostudies-literature
| S-EPMC9131471 | biostudies-literature
| S-EPMC3150427 | biostudies-literature
| S-EPMC10219572 | biostudies-literature
| S-EPMC8781293 | biostudies-literature
| S-EPMC11789145 | biostudies-literature
| S-EPMC3012519 | biostudies-literature
| S-EPMC11570956 | biostudies-literature
| S-EPMC8187880 | biostudies-literature
| S-EPMC8342431 | biostudies-literature