Increasing the Chemical‐Shift Dispersion of Unstructured Proteins with a Covalent Lanthanide Shift Reagent |
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| Authors: | Dr Christoph Göbl Moritz Resch Dr Madeleine Strickland Christoph Hartlmüller Martin Viertler Dr Nico Tjandra Prof Dr Tobias Madl |
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| Institution: | 1. Center for Integrated Protein Science Munich, Technische Universit?t München, Department of Chemistry, Garching, Germany;2. Institute of Structural Biology, Helmholtz Zentrum München, Neuherberg, Germany;3. Laboratory of Structural Biophysics Biochemistry and Biophysics Center, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD, USA;4. Institute of Molecular Biology & Biochemistry, Center of Molecular Medicine, Medical University of Graz, Graz, Austria |
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| 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. |
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| Keywords: | chemical-shift dispersion intrinsically disordered proteins lanthanides NMR pseudo-contact shifts |
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