Cys-Ph-TAHA: a lanthanide binding tag for RDC and PCS enhanced protein NMR
Open Access
- 4 September 2011
- journal article
- research article
- Published by Springer Nature in Journal of Biomolecular NMR
- Vol. 51 (3) , 329-337
Abstract
Here we present Cys-Ph-TAHA, a new nonadentate lanthanide tag for the paramagnetic labelling of proteins. The tag can be easily synthesized and is stereochemically homogenous over a wide range of temperatures, yielding NMR spectra with a single set of peaks. Bound to ubiquitin, it induced large residual dipolar couplings and pseudocontact shifts that could be measured easily and agreed very well with the protein structure. We show that Cys-Ph-TAHA can be used to label large proteins that are biochemically challenging such as the Lac repressor in a 90 kDa ternary complex with DNA and inducer.Keywords
This publication has 50 references indexed in Scilit:
- Weak Long-Range Correlated Motions in a Surface Patch of Ubiquitin Involved in Molecular RecognitionJournal of the American Chemical Society, 2011
- Engineering Encodable Lanthanide-Binding Tags into Loop Regions of ProteinsJournal of the American Chemical Society, 2010
- Flexibility in the Inducer Binding Region Is Crucial for Allostery in the Escherichia coli Lactose RepressorBiochemistry, 2009
- Protein production by auto-induction in high-density shaking culturesProtein Expression and Purification, 2005
- Residual dipolar coupling constants: An elementary derivation of key equationsConcepts in Magnetic Resonance Part A, 2004
- De novo determination of protein structure by NMR using orientational and long-range order restraintsJournal of Molecular Biology, 2000
- Measurement ofJand Dipolar Couplings from Simplified Two-Dimensional NMR SpectraJournal of Magnetic Resonance, 1998
- De novo design of the hydrophobic core of ubiquitinProtein Science, 1997
- NMRPipe: A multidimensional spectral processing system based on UNIX pipesJournal of Biomolecular NMR, 1995
- Temperature dependence of the pseudocontact shift in lanthanide shift reagentsJournal of Magnetic Resonance (1969), 1979