Pseudocontact shifts (PCS) encode long-range information on 3D structures of protein backbones and side-chains. The level of structural detail that can be obtained increases with the number of different sites tagged with a paramagnetic metal ion to generate PCSs. Here we show that PCSs from two different sites can suffice to determine the structure of polypeptide chains and their location and orientation relative to the magnetic susceptibility tensor Ï?, provided that PCSs are available for 1H as well as heteronuclear spins. In addition, PCSs from two different sites are shown to provide detailed structural information on the conformation of methyl group-bearing amino-acid side-chains. A previously published ensemble structure of ubiquitin is shown to explain the magnetic susceptibility and alignment tensors slightly better than structures that try to explain the experimental data by a single conformation, illustrating the potential of PCSs as a tool to investigate small conformational changes.
[NMR paper] Pseudocontact shifts in biomolecular NMR using paramagnetic metal tags.
Pseudocontact shifts in biomolecular NMR using paramagnetic metal tags.
Related Articles Pseudocontact shifts in biomolecular NMR using paramagnetic metal tags.
Prog Nucl Magn Reson Spectrosc. 2017 Feb;98-99:20-49
Authors: Nitsche C, Otting G
PMID: 28283085
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03-12-2017 12:32 PM
Sequence-specific assignment of methyl groups from the neuronal SNARE complex using lanthanide-induced pseudocontact shifts
Sequence-specific assignment of methyl groups from the neuronal SNARE complex using lanthanide-induced pseudocontact shifts
Abstract
Neurotransmitter release depends critically on the neuronal SNARE complex formed by syntaxin-1, SNAP-25 and synaptobrevin, as well as on other proteins such as Munc18-1, Munc13-1 and synaptotagmin-1. Although three-dimensional structures are available for these components, it is still unclear how they are assembled between the synaptic vesicle and plasma membranes to trigger fast, Ca2+-dependent membrane fusion. Methyl...
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12-18-2016 05:04 AM
Pseudocontact shifts in biomolecular NMR using paramagnetic metal tags
Pseudocontact shifts in biomolecular NMR using paramagnetic metal tags
Publication date: Available online 1 December 2016
Source:Progress in Nuclear Magnetic Resonance Spectroscopy</br>
Author(s): Christoph Nitsche, Gottfried Otting</br>
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12-02-2016 07:45 AM
A protocol for the refinement of NMR structures using simultaneously pseudocontact shift restraints from multiple lanthanide ions
A protocol for the refinement of NMR structures using simultaneously pseudocontact shift restraints from multiple lanthanide ions
Abstract
The binding of paramagnetic metal ions to proteins produces a number of different effects on the NMR spectra of the system. In particular, when the magnetic susceptibility of the metal ion is anisotropic, pseudocontact shifts (PCSs) arise and can be easily measured. They constitute very useful restraints for the solution structure determination of metal-binding proteins. In this context, there has been great...
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11-19-2016 08:35 PM
An encodable lanthanide binding tag with reduced size and flexibility for measuring residual dipolar couplings and pseudocontact shifts in large proteins
An encodable lanthanide binding tag with reduced size and flexibility for measuring residual dipolar couplings and pseudocontact shifts in large proteins
Abstract
Metal ions serve important roles in structural biology applications from long-range perturbations seen in magnetic resonance experiments to electron-dense signatures in X-ray crystallography data; however, the metal ion must be secured in a molecular framework to achieve the maximum benefit. Polypeptide-based lanthanide-binding tags (LBTs) represent one option that can be directly encoded...
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01-04-2016 07:49 PM
Using Pseudocontact Shifts and Residual Dipolar Couplingsas Exact NMR Restraints for the Determination of Protein StructuralEnsembles
Using Pseudocontact Shifts and Residual Dipolar Couplingsas Exact NMR Restraints for the Determination of Protein StructuralEnsembles
http://pubs.acs.org/appl/literatum/publisher/achs/journals/content/bichaw/0/bichaw.ahead-of-print/acs.biochem.5b01138/20151217/images/medium/bi-2015-01138j_0006.gif
Biochemistry
DOI: 10.1021/acs.biochem.5b01138
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12-18-2015 07:25 AM
Pulse EPR-enabled interpretation of scarce pseudocontact shifts induced by lanthanide binding tags
Pulse EPR-enabled interpretation of scarce pseudocontact shifts induced by lanthanide binding tags
Abstract
Pseudocontact shifts (PCS) induced by tags loaded with paramagnetic lanthanide ions provide powerful long-range structure information, provided the location of the metal ion relative to the target protein is known. Usually, the metal position is determined by fitting the magnetic susceptibility anisotropy (Î?Ï?) tensor to the 3D structure of the protein in an 8-parameter fit, which requires a large set of PCSs to be reliable. In an alternative...
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11-23-2015 06:58 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...
Structure restraints from heteronuclear pseudocontact shifts generated by lanthanide tags at two different sites
Linear Mode
