title: Bioorthogonally Functionalized NAD(+) Analogues for In-Cell Visualization of Poly(ADP-Ribose) Formation.
abstract: Poly(ADP-ribos)ylation (PARylation) is a major posttranslational modification and signaling event in most eukaryotes. Fundamental processes like DNA repair and transcription are coordinated by this transient polymer and its binding to proteins. ADP-ribosyltransferases (ARTs) build complex ADP-ribose chains from NAD(+) onto various acceptor proteins. Molecular studies of PARylation thus remain challenging. Herein, we present the development of bioorthogonally functionalized NAD(+) analogues for the imaging of PARylation in vitro and in cells. Our results show that 2-modified NAD(+) analogues perform remarkably well and can be applied to the in-cell visualization of PARylation simultaneously in two colors. This tool gives insight into the substrate scope of ARTs and will help to further elucidate the biological role of PARylation by offering fast optical, multichannel read-outs.
authors.affiliation: Department of Chemistry, University of Konstanz, Universitätsstraße 10, 78457, Konstanz, Germany.
authors.forename: Sarah
authors.initials: S
authors.lastname: Wallrodt
authors.affiliation: Department of Chemistry, University of Konstanz, Universitätsstraße 10, 78457, Konstanz, Germany.
authors.forename: Annette
authors.initials: A
authors.lastname: Buntz
authors.affiliation: Department of Chemistry, University of Konstanz, Universitätsstraße 10, 78457, Konstanz, Germany.
authors.forename: Yan
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Department of Chemistry, University of Konstanz, Universitätsstraße 10, 78457, Konstanz, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Zumbusch
authors.affiliation: Department of Chemistry, University of Konstanz, Universitätsstraße 10, 78457, Konstanz, Germany. andreas.marx@uni-konstanz.de.
authors.forename: Andreas
authors.initials: A
authors.lastname: Marx
mesh_terms: D036002:ADP Ribose Transferases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D006861:Hydrogen Peroxide / Q000737:chemistry / Q000378:metabolism; D018613:Microscopy, Confocal; D009243:NAD / Q000031:analogs & derivatives* / Q000378:metabolism; D011064:Poly Adenosine Diphosphate Ribose / Q000737:chemistry*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000378:metabolism
chemical_list: D034741:RNA, Small Interfering; D009243:NAD; D011064:Poly Adenosine Diphosphate Ribose; D006861:Hydrogen Peroxide; D036002:ADP Ribose Transferases
keywords: ADP-ribosylation; bioorthogonal reactions; poly(ADP-ribose); protein modification