Related ArticlesSolid-state NMR Spectra of lipid-anchored Proteins under Magic Angle Spinning.
J Phys Chem B. 2014 Feb 11;
Authors: Nomura K, Harada E, Sugase K, Shimamoto K
Abstract
Solid-state NMR is a promising tool for elucidating membrane-related biological phenomena. We achieved the measurement of high-resolution solid-state NMR spectra for a lipid-anchored protein embedded in lipid bilayers under magic angle spinning (MAS). To date, solid-state NMR measurements of lipid-anchored proteins have not been accomplished due to the difficulty in supplying sufficient amount of stable isotope labeled samples in the overexpression of lipid-anchored proteins requiring complex posttranslational modification. We designed a pseudo-lipid-anchored protein in which the protein component was expressed in E. coli and attached to a chemically synthesized lipid-anchor mimic. Using two types of membranes, liposomes and bicelles, we demonstrated different types of insertion procedures for lipid-anchored protein into membranes. In the liposome sample, we were able to observe the cross polarization and the (13)C-(13)C chemical shift correlation spectra under MAS, indicating that the liposome sample can be used to analyze molecular interactions using dipolar-based NMR experiments. In contrast, the bicelle sample showed sufficient quality of spectra through scalar-based experiments. The relaxation times and protein-membrane interaction were capable of being analyzed in the bicelle sample. These results demonstrated the applicability of two types of sample system to elucidate the roles of lipid-anchors in regulating diverse biological phenomena.
PMID: 24517164 [PubMed - as supplied by publisher]
Shortening spin-lattice relaxation using a copper-chelated lipid at low-temperatures - A magic angle spinning solid-state NMR study on a membrane-bound protein
From The DNP-NMR Blog:
Shortening spin-lattice relaxation using a copper-chelated lipid at low-temperatures - A magic angle spinning solid-state NMR study on a membrane-bound protein
This article is not about DNP. However, the authors describe how to use paramagnetic relaxation enhancers to speed up the data acquisition and with this increase the sensitivity. A similar effect happens when a paramagnetic polarization agent is used in a DNP-NMR experiment and often the only reason why it is actually possible to run 1H-DNP-NMR experiments with recycling delays of several seconds...
[NMR paper] Shortening spin-lattice relaxation using a copper-chelated lipid at low-temperatures - A magic angle spinning solid-state NMR study on a membrane-bound protein.
Shortening spin-lattice relaxation using a copper-chelated lipid at low-temperatures - A magic angle spinning solid-state NMR study on a membrane-bound protein.
Related Articles Shortening spin-lattice relaxation using a copper-chelated lipid at low-temperatures - A magic angle spinning solid-state NMR study on a membrane-bound protein.
J Magn Reson. 2013 Nov 1;237C:175-181
Authors: Yamamoto K, Caporini MA, Im S, Waskell L, Ramamoorthy A
Abstract
Inherent low sensitivity of NMR spectroscopy has been a major disadvantage, especially to...
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11-20-2013 12:52 PM
[NMR paper] Shortening Spin-lattice Relaxation Using a Copper-Chelated lipid at Low-Temperatures – A Magic Angle Spinning Solid-State NMR Study on a Membrane-Bound Protein
Shortening Spin-lattice Relaxation Using a Copper-Chelated lipid at Low-Temperatures – A Magic Angle Spinning Solid-State NMR Study on a Membrane-Bound Protein
Publication date: Available online 1 November 2013
Source:Journal of Magnetic Resonance</br>
Author(s): Kazutoshi Yamamoto , Marc Caporini , Sangchoul Im , Lucy Waskell , Ayyalusamy Ramamoorthy</br>
Inherent low sensitivity of NMR spectroscopy has been a major disadvantage, especially to study biomolecules like membrane proteins. Recent studies have successfully demonstrated the advantages of performing...
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11-01-2013 03:48 AM
[NMR paper] Lipid bilayer preparations of membrane proteins for oriented and magic-angle spinning solid-state NMR samples.
Lipid bilayer preparations of membrane proteins for oriented and magic-angle spinning solid-state NMR samples.
Related Articles Lipid bilayer preparations of membrane proteins for oriented and magic-angle spinning solid-state NMR samples.
Nat Protoc. 2013 Nov;8(11):2256-70
Authors: Das N, Murray DT, Cross TA
Abstract
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10-27-2013 12:53 AM
[NMR paper] Experiments Optimized for Magic Angle Spinning and Oriented Sample Solid-State NMR of Proteins.
Experiments Optimized for Magic Angle Spinning and Oriented Sample Solid-State NMR of Proteins.
Related Articles Experiments Optimized for Magic Angle Spinning and Oriented Sample Solid-State NMR of Proteins.
J Phys Chem B. 2013 Sep 17;
Authors: Das BB, Lin EC, Opella SJ
Abstract
Structure determination by solid-state NMR of proteins is rapidly advancing as result of recent developments of samples, experimental methods, and calculations. There are a number of different solid-state NMR approaches that utilize stationary, aligned samples or...
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09-21-2013 06:50 PM
[NMR paper] Orphan spin operators enable the acquisition of multiple 2D and 3D magic angle spinning solid-state NMR spectra.
Orphan spin operators enable the acquisition of multiple 2D and 3D magic angle spinning solid-state NMR spectra.
Related Articles Orphan spin operators enable the acquisition of multiple 2D and 3D magic angle spinning solid-state NMR spectra.
J Chem Phys. 2013 May 14;138(18):184201
Authors: Gopinath T, Veglia G
Abstract
We propose a general method that enables the acquisition of multiple 2D and 3D solid-state NMR spectra for U-(13)C, (15)N-labeled proteins. This method, called MEIOSIS (Multiple ExperIments via Orphan SpIn operatorS), makes...
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05-17-2013 07:00 PM
Solid-state magic-angle spinning NMR of membrane proteins and protein–ligand interactions
Solid-state magic-angle spinning NMR of membrane proteins and protein–ligand interactions
April 2012
Publication year: 2012
Source:European Journal of Cell Biology, Volume 91, Issue 4</br>
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Structural biology is developing into a universal tool for visualizing biological processes in space and time at atomic resolution. The field has been built by established methodology like X-ray crystallography, electron microscopy and solution NMR and is now incorporating new techniques, such as small-angle X-ray scattering, electron tomography, magic-angle-spinning solid-state...