Journal
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
Volume 55, Issue 47, Pages 14847-14851Publisher
WILEY-V C H VERLAG GMBH
DOI: 10.1002/anie.201607261
Keywords
chemical-shift dispersion; intrinsically disordered proteins; lanthanides; NMR; pseudo-contact shifts
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Funding
- Intramural Research Program of the NIH, NHLBI
- Bavarian Ministry of Sciences, Research and the Arts (Bavarian Molecular Biosystems Research Network)
- German Research Foundation (Emmy Noether program) [MA 5703/1-1]
- Center for Integrated Protein Science Munich (CIPSM)
- Austrian Science Fund [FWF: P28854, W1226-B18]
- Austrian Science Fund (FWF) [P28854] Funding Source: Austrian Science Fund (FWF)
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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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