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 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 non-specific 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, (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.
PMID: 21877751 [PubMed - as supplied by publisher]
[Question from NMRWiki Q&A forum] Spectra from BMRB Chemical shifts
Spectra from BMRB Chemical shifts
Is it possible to generate a spectrum from BMRB chemical shifts?
I want to generate a spectrum to use as a reference for an H1-N15 HSQC experiment.
Check if somebody has answered this question on NMRWiki QA forum
nmrlearner
News from other NMR forums
0
11-04-2011 03:17 AM
[NMR images] Chemical Shifts in NMR Spectra
http://hyperphysics.phy-astr.gsu.edu/hbase/nuclear/imgnuc/cshift.gif
hyperphysics.phy-astr.gsu.edu
5/07/2011 6:04:05 PM GMT
Chemical Shifts in NMR Spectra
More...
nmrlearner
NMR pictures
0
07-26-2011 11:22 PM
Proteinā??protein HADDocking using exclusively pseudocontact shifts
Proteinā??protein HADDocking using exclusively pseudocontact shifts
<div class="Abstract" lang="en">Abstract <div class="normal">In order to enhance the structure determination process of macromolecular assemblies by NMR, we have implemented long-range pseudocontact shift (PCS) restraints into the data-driven protein docking package HADDOCK. We demonstrate the efficiency of the method on a synthetic, yet realistic case based on the lanthanide-labeled N-terminal Īµ domain of the E. coli DNA polymerase III (Īµ186) in complex with the HOT domain. Docking from the bound form of the two...
nmrlearner
Journal club
0
06-06-2011 12:53 AM
Generation of Pseudocontact Shifts in Protein NMR Spectra with a Genetically Encoded Cobalt(II)-Binding Amino Acid.
Generation of Pseudocontact Shifts in Protein NMR Spectra with a Genetically Encoded Cobalt(II)-Binding Amino Acid.
Generation of Pseudocontact Shifts in Protein NMR Spectra with a Genetically Encoded Cobalt(II)-Binding Amino Acid.
Angew Chem Int Ed Engl. 2011 Jan 17;50(3):692-4
Authors: Nguyen TH, Ozawa K, Stanton-Cook M, Barrow R, Huber T, Otting G
nmrlearner
Journal club
0
01-13-2011 12:00 PM
Generation of Pseudocontact Shifts in Protein NMR Spectra with a Genetically Encoded
Generation of Pseudocontact Shifts in Protein NMR Spectra with a Genetically Encoded Cobalt(II)-Binding Amino Acid.
Related Articles Generation of Pseudocontact Shifts in Protein NMR Spectra with a Genetically Encoded Cobalt(II)-Binding Amino Acid.
Angew Chem Int Ed Engl. 2010 Nov 25;
Authors: Nguyen TH, Ozawa K, Stanton-Cook M, Barrow R, Huber T, Otting G
nmrlearner
Journal club
0
11-27-2010 02:45 PM
[NMR paper] More reliable protein NMR peak assignment via improved 2-interval scheduling.
More reliable protein NMR peak assignment via improved 2-interval scheduling.
Related Articles More reliable protein NMR peak assignment via improved 2-interval scheduling.
J Comput Biol. 2005 Mar;12(2):129-46
Authors: Chen ZZ, Lin G, Rizzi R, Wen J, Xu D, Xu Y, Jiang T
Protein NMR peak assignment refers to the process of assigning a group of "spin systems" obtained experimentally to a protein sequence of amino acids. The automation of this process is still an unsolved and challenging problem in NMR protein structure determination. Recently,...
nmrlearner
Journal club
0
11-24-2010 11:14 PM
[NMR paper] Improving the accuracy of NMR structures of large proteins using pseudocontact shifts
Improving the accuracy of NMR structures of large proteins using pseudocontact shifts as long-range restraints.
Related Articles Improving the accuracy of NMR structures of large proteins using pseudocontact shifts as long-range restraints.
J Biomol NMR. 2004 Mar;28(3):205-12
Authors: Gaponenko V, Sarma SP, Altieri AS, Horita DA, Li J, Byrd RA
We demonstrate improved accuracy in protein structure determination for large (>/=30 kDa), deuterated proteins (e.g. STAT4(NT)) via the combination of pseudocontact shifts for amide and methyl protons...
nmrlearner
Journal club
0
11-24-2010 09:25 PM
Numbat: an interactive software tool for fitting Δχ-tensors to molecular coordinates using pseudocontact shifts
Numbat: an interactive software tool for fitting Δχ-tensors to molecular coordinates using pseudocontact shifts
Christophe Schmitz, Mitchell J. Stanton-Cook, Xun-Cheng Su, Gottfried Otting and Thomas Huber
Journal of Biomolecular NMR; 2008; 41(3) pp 179 - 189
Abstract:
Pseudocontact shift (PCS) effects induced by a paramagnetic lanthanide bound to a protein have become increasingly popular in NMR spectroscopy as they yield a complementary set of orientational and long-range structural restraints. PCS are a manifestation of the χ-tensor anisotropy, the Δχ-tensor, which in turn can be...
A DOTA-amide lanthanide tag for reliable generation of pseudocontact shifts in protein NMR spectra.
Linear Mode
