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.
Content Type Journal Article
Category Article
Pages 1-9
DOI 10.1007/s10858-011-9560-y
Authors
Fabian Peters, Department of NMR-Based Structural Biology, Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, 37077 Göttingen, Germany
Mitcheell Maestre-Martinez, Department of NMR-Based Structural Biology, Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, 37077 Göttingen, Germany
Andrei Leonov, Department of NMR-Based Structural Biology, Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, 37077 Göttingen, Germany
Lidija KovaÄ?iÄ?, Bijvoet Center for Biomolecular Research, Department of NMR Spectroscopy, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands
Stefan Becker, Department of NMR-Based Structural Biology, Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, 37077 Göttingen, Germany
Rolf Boelens, Bijvoet Center for Biomolecular Research, Department of NMR Spectroscopy, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands
Christian Griesinger, Department of NMR-Based Structural Biology, Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, 37077 Göttingen, Germany
Cys-Ph-TAHA: a lanthanide binding tag for RDC and PCS enhanced protein NMR.
Cys-Ph-TAHA: a lanthanide binding tag for RDC and PCS enhanced protein NMR.
Cys-Ph-TAHA: a lanthanide binding tag for RDC and PCS enhanced protein NMR.
J Biomol NMR. 2011 Sep 4;
Authors: Peters F, Maestre-Martinez M, Leonov A, Kova?i? L, Becker S, Boelens R, Griesinger C
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....
nmrlearner
Journal club
0
09-06-2011 06:02 PM
A DOTA-amide lanthanide tag for reliable generation of pseudocontact shifts in protein NMR spectra.
A DOTA-amide lanthanide tag for reliable generation of pseudocontact shifts in protein NMR spectra.
A DOTA-amide lanthanide tag for reliable generation of pseudocontact shifts in protein NMR spectra.
Bioconjug Chem. 2011 Aug 31;
Authors: Graham B, Loh CT, Swarbrick JD, Ung P, Shin J, Yagi H, Jia X, Chhabra S, Barlow N, Pintacuda G, Huber T, Otting G
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...
nmrlearner
Journal club
0
09-01-2011 05:20 PM
An iminodiacetic acid based lanthanide binding tag for paramagnetic exchange NMR spectroscopy.
An iminodiacetic acid based lanthanide binding tag for paramagnetic exchange NMR spectroscopy.
An iminodiacetic acid based lanthanide binding tag for paramagnetic exchange NMR spectroscopy.
Angew Chem Int Ed Engl. 2011 May 2;50(19):4403-6
Authors: Swarbrick JD, Ung P, Chhabra S, Graham B
PMID: 21480444
nmrlearner
Journal club
0
08-19-2011 02:56 PM
Engineering [Ln(DPA)3]3â?? binding sites in proteins: a widely applicable method for tagging proteins with lanthanide ions
Engineering 3â?? binding sites in proteins: a widely applicable method for tagging proteins with lanthanide ions
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...
nmrlearner
Journal club
0
07-26-2011 11:11 AM
Engineering of a bis-chelator motif into a protein ?-helix for rigid lanthanide binding and paramagnetic NMR spectroscopy.
Engineering of a bis-chelator motif into a protein ?-helix for rigid lanthanide binding and paramagnetic NMR spectroscopy.
Engineering of a bis-chelator motif into a protein ?-helix for rigid lanthanide binding and paramagnetic NMR spectroscopy.
Chem Commun (Camb). 2011 May 27;
Authors: Swarbrick JD, Ung P, Su XC, Maleckis A, Chhabra S, Huber T, Otting G, Graham B
Attachment of two nitrilotriacetic acid-based ligands to a protein ?-helix in an i, i + 4 configuration produces an octadentate chelating motif that is able to bind paramagnetic...
nmrlearner
Journal club
0
05-28-2011 06:50 PM
A rigid lanthanide binding tag for NMR structural analysis of carbohydrates.
A rigid lanthanide binding tag for NMR structural analysis of carbohydrates.
A rigid lanthanide binding tag for NMR structural analysis of carbohydrates.
Chem Commun (Camb). 2011 May 23;
Authors: Mallagaray A, Canales A, Domínguez G, Jiménez-Barbero J, Pérez-Castells J
The first synthesis of a carbohydrate molecule covalently bound to a rigid lanthanide binding tag is reported. This derivative has been designed as a new tool to provide long-range restraints for structural elucidation and molecular recognition studies of carbohydrates, thus...
nmrlearner
Journal club
0
05-25-2011 07:01 PM
4,4'-Dithiobis-dipicolinic Acid: A Small and Convenient Lanthanide Binding Tag for Protein NMR Spectroscopy.
4,4'-Dithiobis-dipicolinic Acid: A Small and Convenient Lanthanide Binding Tag for Protein NMR Spectroscopy.
4,4'-Dithiobis-dipicolinic Acid: A Small and Convenient Lanthanide Binding Tag for Protein NMR Spectroscopy.
Chemistry. 2011 May 3;
Authors: Jia X, Maleckis A, Huber T, Otting G
Pseudocontact shifts (PCS) from paramagnetic lanthanide ions present powerful long-range structure restraints for studies of proteins by nuclear magnetic resonance spectroscopy. To elicit PCSs, the lanthanide must be attached site-specifically to the target protein....
nmrlearner
Journal club
0
05-06-2011 02:00 AM
[NMR paper] A caged lanthanide complex as a paramagnetic shift agent for protein NMR.
A caged lanthanide complex as a paramagnetic shift agent for protein NMR.
Related Articles A caged lanthanide complex as a paramagnetic shift agent for protein NMR.
Chemistry. 2004 Jul 5;10(13):3252-60
Authors: Prudêncio M, Rohovec J, Peters JA, Tocheva E, Boulanger MJ, Murphy ME, Hupkes HJ, Kosters W, Impagliazzo A, Ubbink M
A lanthanide complex, named CLaNP (caged lanthanide NMR probe) has been developed for the characterisation of proteins by paramagnetic NMR spectroscopy. The probe consists of a lanthanide chelated by a derivative of DTPA...
Cys-Ph-TAHA: a lanthanide binding tag for RDC and PCS enhanced protein NMR
Linear Mode
