title: Increasing the Chemical-Shift Dispersion of Unstructured Proteins with a Covalent Lanthanide Shift Reagent.
abstract: The study of intrinsically disordered proteins (IDPs) by NMR often suffers from highly overlapped resonances that prevent unambiguous chemical-shift assignments, and data analysis that relies on well-separated resonances. We present a covalent paramagnetic lanthanide-binding tag (LBT) for increasing the chemical-shift dispersion and facilitating the chemical-shift assignment of challenging, repeat-containing IDPs. Linkage of the DOTA-based LBT to a cysteine residue induces pseudo-contact shifts (PCS) for resonances more than 20 residues from the spin-labeling site. This leads to increased chemical-shift dispersion and decreased signal overlap, thereby greatly facilitating chemical-shift assignment. This approach is applicable to IDPs of varying sizes and complexity, and is particularly helpful for repeat-containing IDPs and low-complexity regions. This results in improved efficiency for IDP analysis and binding studies.
authors.affiliation: Center for Integrated Protein Science Munich, Technische Universität München, Department of Chemistry, Lichtenbergstraße 4, 85748, Garching, Germany.|Institute of Structural Biology, Helmholtz Zentrum München, Ingolstädter Landstr. 1, 85764, Neuherberg, Germany.
authors.forename: Christoph
authors.identifier: 0000-0001-8614-1434
authors.initials: C
authors.lastname: Göbl
authors.affiliation: Center for Integrated Protein Science Munich, Technische Universität München, Department of Chemistry, Lichtenbergstraße 4, 85748, Garching, Germany.|Institute of Structural Biology, Helmholtz Zentrum München, Ingolstädter Landstr. 1, 85764, Neuherberg, Germany.
authors.forename: Moritz
authors.identifier: 0000-0003-3477-3565
authors.initials: M
authors.lastname: Resch
authors.affiliation: Laboratory of Structural Biophysics Biochemistry and Biophysics Center, National Heart, Lung, and Blood Institute, National Institutes of Health, Building 50, Bethesda, MD, 20814, USA.
authors.forename: Madeleine
authors.identifier: 0000-0001-8160-070X
authors.initials: M
authors.lastname: Strickland
authors.affiliation: Center for Integrated Protein Science Munich, Technische Universität München, Department of Chemistry, Lichtenbergstraße 4, 85748, Garching, Germany.|Institute of Structural Biology, Helmholtz Zentrum München, Ingolstädter Landstr. 1, 85764, Neuherberg, Germany.
authors.forename: Christoph
authors.initials: C
authors.lastname: Hartlmüller
authors.affiliation: Institute of Structural Biology, Helmholtz Zentrum München, Ingolstädter Landstr. 1, 85764, Neuherberg, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Viertler
authors.affiliation: Laboratory of Structural Biophysics Biochemistry and Biophysics Center, National Heart, Lung, and Blood Institute, National Institutes of Health, Building 50, Bethesda, MD, 20814, USA.
authors.forename: Nico
authors.initials: N
authors.lastname: Tjandra
authors.affiliation: Center for Integrated Protein Science Munich, Technische Universität München, Department of Chemistry, Lichtenbergstraße 4, 85748, Garching, Germany.|Institute of Structural Biology, Helmholtz Zentrum München, Ingolstädter Landstr. 1, 85764, Neuherberg, Germany.|Institute of Molecular Biology & Biochemistry, Center of Molecular Medicine, Medical University of Graz, 8010, Graz, Austria.
authors.forename: Tobias
authors.identifier: 0000-0002-9725-5231
authors.initials: T
authors.lastname: Madl
mesh_terms: D064267:Intrinsically Disordered Proteins / Q000737:chemistry*; D028581:Lanthanoid Series Elements / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy
chemical_list: D064267:Intrinsically Disordered Proteins; D028581:Lanthanoid Series Elements
keywords: NMR; chemical-shift dispersion; intrinsically disordered proteins; lanthanides; pseudo-contact shifts