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The photocatalyzed thiol-ene reaction: A new tag to yield fast, selective and reversible paramagnetic tagging of proteins.
ChemPhysChem 21, 863-869 (2020)
Paramagnetic restraints have been used in biomolecular NMR for the last three decades to elucidate and refine biomolecular structures, but also to characterize protein-ligand interactions. A common technique to generate such restraints in proteins, which do not naturally contain a (paramagnetic) metal, consists in the attachment to the protein of a lanthanide-binding-tag (LBT). In order to design such LBTs, it is important to consider the efficiency and stability of the conjugation, the geometry of the complex (conformational exchanges and coordination) and the chemical inertness of the ligand. Here we describe a photo-catalyzed thiol-ene reaction for the cysteine-selective paramagnetic tagging of proteins. As a model, we designed an LBT with a vinyl-pyridine moiety which was used to attach our tag to the protein GB1 in fast and irreversible fashion. Our tag T1 yields magnetic susceptibility tensors of significant size with different lanthanides and has been characterized using NMR and relaxometry measurements.
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Publikationstyp Artikel: Journalartikel
Dokumenttyp Wissenschaftlicher Artikel
Schlagwörter Biomolecular Nmr ; Lanthanide Coordination ; Paramagnetic Tags ; Photocatalyzed Thiol-ene Reaction ; Protein Structure Refinement; Pseudocontact Shifts; X-ray; Nmr; Complexes; Binding; Coordination; Constraints; Inhibitors; Dynamics; Import
ISSN (print) / ISBN 1439-4235
Quellenangaben Band: 21, Heft: 9, Seiten: 863-869
Verlagsort Postfach 101161, 69451 Weinheim, Germany
Begutachtungsstatus Peer reviewed
Institut(e) Institute of Structural Biology (STB)