Design and applications of lanthanide chelating tags for pseudocontact shift NMR spectroscopy with biomacromolecules
In this review, lanthanide chelating tags and their application in pseudocontact shift NMR spectroscopy as well as analysis of residual dipolar couplings are covered. Including a complete overview of non-DOTA-derived and DOTAderived lanthanide chelating tags, critical points in the design of lanthanide chelating tags as appropriate linker moieties, stability under reductive conditions e.g. for in-cell applications, magnitude of the transferred anisotropy from the lanthanide chelating tag to the
... helating tag to the biomacromolecule under investigation and structural properties as well as conformational bias of the lanthanide chelating tags are discussed. Furthermore, all currently published DOTAderived lanthanide chelating tags used for PCS NMR spectroscopy are displayed in tabular form including their anisotropy parameters with all employed lanthanide ions, CB-Ln distances and tagging reaction conditions, i.e. stoichiometry of lanthanide chelating tag, pH, buffer composition, temperature and reaction time. Additionally, reported applications of lanthanide chelating tags for pseudocontact shifts and residual dipolar couplings on proteins, protein-protein and protein-ligand complexes, carbohydrates, carbohydrate-protein complexes, nucleic acids and nucleic acid-protein complexes are presented and critically reviewed. The vast and impressing field of applications of lanthanide chelating tags in structural investigations of biomacromolecules in solution clearly illustrates the significance of this particular field of research and the extension of the repertoire of lanthanide chelating tags from proteins to nucleic acids holds great promise for the determination of valuable structural parameters and further developments in characterising intermolecular interactions.