title: New AdoMet Analogues as Tools for Enzymatic Transfer of Photo-Cross-Linkers and Capturing RNA-Protein Interactions.
abstract: Elucidation of biomolecular interactions is of utmost importance in biochemistry. Photo-cross-linking offers the possibility to precisely determine RNA-protein interactions. However, despite the inherent specificity of enzymes, approaches for site-specific introduction of photo-cross-linking moieties into nucleic acids are scarce. Methyltransferases in combination with synthetic analogues of their natural cosubstrate S-adenosyl-l-methionine (AdoMet) allow for the post-synthetic site-specific modification of biomolecules. We report on three novel AdoMet analogues bearing the most widespread photo-cross-linking moieties (aryl azide, diazirine, and benzophenone). We show that these photo-cross-linkers can be enzymatically transferred to the methyltransferase target, that is, the mRNA cap, with high efficiency. Photo-cross-linking of the resulting modified mRNAs with the cap interacting protein eIF4E was successful with aryl azide and diazirine but not benzophenone, reflecting the affinity of the modified 5' caps.
authors.affiliation: University of Münster, Department of Chemistry, Institute of Biochemistry, Wilhelm-Klemm-Str. 2, 48149, Münster, Germany.
authors.forename: Fabian
authors.initials: F
authors.lastname: Muttach
authors.affiliation: University of Münster, Department of Chemistry, Institute of Organic Chemistry, Corrensstr. 40, 48149, Münster, Germany.
authors.forename: Florian
authors.initials: F
authors.lastname: Mäsing
authors.affiliation: University of Münster, Department of Chemistry, Institute of Organic Chemistry, Corrensstr. 40, 48149, Münster, Germany.
authors.forename: Armido
authors.identifier: http://orcid.org/0000-0002-1706-513X
authors.initials: A
authors.lastname: Studer
authors.affiliation: University of Münster, Department of Chemistry, Institute of Biochemistry, Wilhelm-Klemm-Str. 2, 48149, Münster, Germany.|Cells-in-Motion Cluster of Excellence (EXC 1003-CiM), University of Münster, Germany.
authors.forename: Andrea
authors.initials: A
authors.lastname: Rentmeister
keywords: AdoMet analogue; RNA labelling; RNA recognition; RNA-protein interactions; photo-cross-linking