Details

Autor(en) / Beteiligte

• Göbl, Christoph
• Resch, Moritz
• Strickland, Madeleine
• Hartlmüller, Christoph
• Viertler, Martin
• Tjandra, Nico
• Madl, Tobias

Titel

Increasing the Chemical-Shift Dispersion of Unstructured Proteins with a Covalent Lanthanide Shift Reagent

Ist Teil von

• Angewandte Chemie (International ed.), 2016-11, Vol.55 (47), p.14847-14851

Auflage

International ed. in English

Ort / Verlag

Germany: Blackwell Publishing Ltd

Erscheinungsjahr

2016

Quelle

MEDLINE

Beschreibungen/Notizen

• 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. Widespread shift: A covalent paramagnetic lanthanide‐binding tag (LBT) was used to increase the chemical‐shift dispersion in protein NMR spectroscopy, thus facilitating the chemical‐shift assignment of challenging intrinsically disordered proteins. 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 (DOTA=1,4,7,10‐tetraazacyclododecane‐1,4,7,10‐tetraacetic acid).