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

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...

Full description

Saved in:
Bibliographic Details
Published in:Angewandte Chemie International Edition 2016-11, Vol.55 (47), p.14847-14851
Main Authors: Göbl, Christoph, Resch, Moritz, Strickland, Madeleine, Hartlmüller, Christoph, Viertler, Martin, Tjandra, Nico, Madl, Tobias
Format: Article
Language:English
Subjects:
Binding
chemical-shift dispersion
Communication
Communications
Complexity
Covalence
Cysteine
Data analysis
Data processing
Dispersion
intrinsically disordered proteins
Intrinsically Disordered Proteins - chemistry
lanthanides
Lanthanoid Series Elements - chemistry
Magnetic Resonance Spectroscopy
NMR
Nuclear magnetic resonance
Proteins
pseudo-contact shifts
Spin labeling
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary: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).
ISSN:1433-7851
1521-3773
1521-3773
DOI:10.1002/anie.201607261