title: A Biocatalytic Cascade for Versatile One-Pot Modification of mRNA Starting from Methionine Analogues.
abstract: 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.
authors.affiliation: University of Münster, Department of Chemistry, Institute of Biochemistry, Wilhelm-Klemm-Strasse 2, 48149, Münster, Germany.
authors.forename: Fabian
authors.initials: F
authors.lastname: Muttach
authors.affiliation: University of Münster, Department of Chemistry, Institute of Biochemistry, Wilhelm-Klemm-Strasse 2, 48149, Münster, Germany. a.rentmeister@uni-muenster.de.|Cells-in-Motion Cluster of Excellence (EXC 1003 - CiM), University of Münster, Germany. a.rentmeister@uni-muenster.de.
authors.forename: Andrea
authors.initials: A
authors.lastname: Rentmeister
mesh_terms: D055162:Biocatalysis; D008715:Methionine / Q000737:chemistry*; D012333:RNA, Messenger / Q000737:chemistry*
chemical_list: D012333:RNA, Messenger; D008715:Methionine
keywords: 5′ cap; RNA; SAM synthetase; click chemistry; labeling