Engineering [Ln(DPA)3]3- binding sites in proteins: a widely applicable method for tagging proteins with lanthanide ions

Jia, Xinying, Yagi, Hiromasa, Su, Xun-Cheng, Stanton-Cook, Mitchell, Huber, Thomas and Otting, Gottfried (2011) Engineering [Ln(DPA)3]3- binding sites in proteins: a widely applicable method for tagging proteins with lanthanide ions. Journal of Biomolecular NMR, 50 411-420. doi:10.1007/s10858-011-9529-x


Author Jia, Xinying
Yagi, Hiromasa
Su, Xun-Cheng
Stanton-Cook, Mitchell
Huber, Thomas
Otting, Gottfried
Title Engineering [Ln(DPA)3]3- binding sites in proteins: a widely applicable method for tagging proteins with lanthanide ions
Formatted title
Engineering [Ln(DPA)3]3- binding sites in proteins: a widely applicable method for tagging proteins with lanthanide ions
Journal name Journal of Biomolecular NMR   Check publisher's open access policy
ISSN 0925-2738
1573-5001
Publication date 2011-08
Year available 2011
Sub-type Article (original research)
DOI 10.1007/s10858-011-9529-x
Open Access Status Not yet assessed
Volume 50
Start page 411
End page 420
Total pages 10
Place of publication Dordrecht, Netherlands
Publisher Springer Netherlands
Language eng
Formatted abstract
Paramagnetic relaxation enhancements from unpaired electrons observed in nuclear magnetic resonance (NMR) spectra present powerful long-range distance restraints. The most frequently used paramagnetic tags, however, are tethered to the protein via disulfide bonds, requiring proteins with single cysteine residues for covalent attachment. Here we present a straightforward strategy to tag proteins site-specifically with paramagnetic lanthanides without a tether and independent of cysteine residues. It relies on preferential binding of the complex between three dipicolinic acid molecules (DPA) and a lanthanide ion (Ln3+), [Ln(DPA3]3-, to a pair of positively charged amino acids whose charges are not compensated by negatively charged residues nearby. This situation rarely occurs in wild-type proteins, allowing the creation of specific binding sites simply by introduction of positively charged residues that are positioned far from glutamate or aspartate residues. The concept is demonstrated with the hnRNPLL RRM1 domain. In addition, we show that histidine- and arginine-tags present binding sites for [Ln(DPA)3]3-.
Keyword [Ln(DPA)3]3-
Lanthanide tag
NMR spectroscopy
Paramagnetic relaxation enhancements
RRM domain
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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