期刊
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
卷 55, 期 5, 页码 1917-1920出版社
WILEY-V C H VERLAG GMBH
DOI: 10.1002/anie.201507577
关键词
5 ' cap; click chemistry; labeling; RNA; SAM synthetase
资金
- DFG [RE 2796/2-1]
- Fonds der Chemischen Industrie
- DFG Priority Programme SPP 1784 [RE 2796/3-1]
Methyltransferases have proven useful to install functional groups site-specifically in different classes of biomolecules when analogues of their cosubstrate S-adenosyl-L-methionine (AdoMet) are available. Methyltransferases have been used to address different classes of RNA molecules selectively and site-specifically, which is indispensable for biophysical and mechanistic studies as well as labeling in the complex cellular environment. However, the AdoMet analogues are not cell-permeable, thus preventing implementation of this strategy in cells. We present a two-step enzymatic cascade for site-specific mRNA modification starting from stable methionine analogues. Our approach combines the enzymatic synthesis of AdoMet with modification of the 5' cap by a specific RNA methyltransferase in one pot. We demonstrate that a substrate panel including alkene, alkyne, and azido functionalities can be used and further derivatized in different types of click reactions.
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