DOTA-amide lanthanide tag for reliable generation of pseudocontact shifts in protein NMR spectra

Graham, Bim, Loh, Choy Theng, Swarbrick, James David, Ung, Phuc, Shin, James, Yagi, Hiromasa, Jia, Xinying, Chhabra, Sandeep, Barlow, Nicholas, Pintacuda, Guido, Huber, Thomas and Otting, Gottfried (2011) DOTA-amide lanthanide tag for reliable generation of pseudocontact shifts in protein NMR spectra. Bioconjugate Chemistry, 22 10: 2118-2125. doi:10.1021/bc200353c

Author Graham, Bim
Loh, Choy Theng
Swarbrick, James David
Ung, Phuc
Shin, James
Yagi, Hiromasa
Jia, Xinying
Chhabra, Sandeep
Barlow, Nicholas
Pintacuda, Guido
Huber, Thomas
Otting, Gottfried
Title DOTA-amide lanthanide tag for reliable generation of pseudocontact shifts in protein NMR spectra
Journal name Bioconjugate Chemistry   Check publisher's open access policy
ISSN 1043-1802
Publication date 2011-10-19
Year available 2011
Sub-type Article (original research)
DOI 10.1021/bc200353c
Open Access Status Not yet assessed
Volume 22
Issue 10
Start page 2118
End page 2125
Total pages 8
Place of publication Washington, DC, United States
Publisher American Chemical Society
Language eng
Formatted abstract
Structural studies of proteins and protein-ligand complexes by nuclear magnetic resonance (NMR) spectroscopy can be greatly enhanced by site-specific attachment of lanthanide ions to create paramagnetic centers. In particular, pseudocontact shifts (PCS) generated by paramagnetic lanthanides contain important and unique long-range structure information. Here, we present a high-affinity lanthanide binding tag that can be attached to single cysteine residues of proteins. The new tag has many advantageous features that are not available in this combination from previously published tags: (i) it binds lanthanide ions very tightly, minimizing the generation of nonspecific effects, (ii) it produces PCSs with high reliability as its bulkiness prevents complete motional averaging of PCSs, (iii) it can be attached to single cysteine residues, alleviating the need of detailed prior knowledge of the 3D structure of the target protein, and (iv) it does not display conformational exchange phenomena that would increase the number of signals in the NMR spectrum. The performance of the tag is demonstrated with the N-terminal domain of the E. coli arginine repressor and the A28C mutant of human ubiquitin.
Keyword Biochemical Research Methods
Biochemistry & Molecular Biology
Chemistry, Multidisciplinary
Chemistry, Organic
Biochemistry & Molecular Biology
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status Non-UQ

Document type: Journal Article
Sub-type: Article (original research)
Collection: School of Chemistry and Molecular Biosciences
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Citation counts: TR Web of Science Citation Count  Cited 60 times in Thomson Reuters Web of Science Article | Citations
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