Publication date: Available online 22 December 2017 Source:Methods
Author(s): Songlin Wang, T. Gopinath, Gianluigi Veglia
Oriented sample solid-state NMR (OS-ssNMR) spectroscopy is uniquely suited to determine membrane protein topology at the atomic resolution in liquid crystalline bilayers under physiological temperature. However, the inherent low sensitivity of this technique has hindered the throughput of multidimensional experiments necessary for resonance assignments and structure determination. In this work, we show that doping membrane protein bicelle preparations with paramagnetic ion chelated lipids and exploiting paramagnetic relaxation effects it is possible to accelerate the acquisition of both 2D and 3D multidimensional experiments with significant saving in time. We demonstrate the efficacy of this method for a small membrane protein, sarcolipin, reconstituted in DMPC/POPC/DHPC oriented bicelles. In particular, using Cu2+-DMPE-DTPA as a dopant, we observed a decrease of 1H T1 of sarcolipin by 2/3, allowing us to reduce the recycle delay up to 3 times. We anticipate that these new developments will enable the routine acquisition of multidimensional OS-ssNMR experiments.
[NMR paper] Rapid Quantitative Measurements of Paramagnetic Relaxation Enhancements in Cu(II)-Tagged Proteins by Proton-Detected Solid-State NMR Spectroscopy.
Rapid Quantitative Measurements of Paramagnetic Relaxation Enhancements in Cu(II)-Tagged Proteins by Proton-Detected Solid-State NMR Spectroscopy.
Related Articles Rapid Quantitative Measurements of Paramagnetic Relaxation Enhancements in Cu(II)-Tagged Proteins by Proton-Detected Solid-State NMR Spectroscopy.
J Phys Chem Lett. 2017 Nov 17;:
Authors: Mukhopadhyay D, Nadaud PS, Shannon MD, Jaroniec CP
Abstract
We demonstrate rapid quantitative measurements of site-resolved paramagnetic relaxation enhancements (PREs), which are a...
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[NMR paper] Characterization of protein-protein interfaces in large complexes by solid state NMR solvent paramagnetic relaxation enhancements.
Characterization of protein-protein interfaces in large complexes by solid state NMR solvent paramagnetic relaxation enhancements.
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J Am Chem Soc. 2017 Aug 07;:
Authors: Öster C, Kosol S, Hartlmüller C, Lamley JM, Iuga D, Oss A, Org ML, Vanatalu K, Samoson A, Madl T, Lewandowski JR
Abstract
Solid-state NMR is becoming a viable alternative for obtaining information about structures and...
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[NMR paper] Simultaneous acquisition of 2D and 3D solid-state NMR experiments for sequential assignment of oriented membrane protein samples.
Simultaneous acquisition of 2D and 3D solid-state NMR experiments for sequential assignment of oriented membrane protein samples.
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J Biomol NMR. 2015 Mar 7;
Authors: Gopinath T, Mote KR, Veglia G
Abstract
We present a new method called DAISY (Dual Acquisition orIented ssNMR spectroScopY) for the simultaneous acquisition of 2D and 3D oriented solid-state NMR experiments for membrane...
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03-10-2015 07:22 PM
Simultaneous acquisition of 2D and 3D solid-state NMR experiments for sequential assignment of oriented membrane protein samples
Simultaneous acquisition of 2D and 3D solid-state NMR experiments for sequential assignment of oriented membrane protein samples
Abstract
We present a new method called DAISY (Dual Acquisition orIented ssNMR spectroScopY) for the simultaneous acquisition of 2D and 3D oriented solid-state NMR experiments for membrane proteins reconstituted in mechanically or magnetically aligned lipid bilayers. DAISY utilizes dual acquisition of sine and cosine dipolar or chemical shift coherences and long living 15N longitudinal polarization to obtain two...
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[NMR paper] Sensitivity and resolution enhancement of oriented solid-state NMR: Application to membrane proteins.
Sensitivity and resolution enhancement of oriented solid-state NMR: Application to membrane proteins.
Related Articles Sensitivity and resolution enhancement of oriented solid-state NMR: Application to membrane proteins.
Prog Nucl Magn Reson Spectrosc. 2013 Nov;75:50-68
Authors: Gopinath T, Mote KR, Veglia G
Abstract
Oriented solid-state NMR (O-ssNMR) spectroscopy is a major technique for the high-resolution analysis of the structure and topology of transmembrane proteins in native-like environments. Unlike magic angle spinning (MAS)...
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Sensitivity and Resolution Enhancement of Oriented Solid-State NMR: Application to Membrane Proteins
Sensitivity and Resolution Enhancement of Oriented Solid-State NMR: Application to Membrane Proteins
Publication date: Available online 12 August 2013
Source:Progress in Nuclear Magnetic Resonance Spectroscopy</br>
Author(s): T. Gopinath , Kaustubh R. Mote , Gianluigi Veglia</br>
Oriented solid-state NMR (O-ssNMR) spectroscopy is a major technique for the high-resolution analysis of the structure and topology of transmembrane proteins in native-like environments. Unlike magic angle spinning (MAS) techniques, O-ssNMR spectroscopy requires membrane protein...