Related Articles1H/13C/15N triple-resonance experiments for structure determinaton of membrane proteins by oriented-sample NMR.
Solid State Nucl Magn Reson. 2020 Nov 24;111:101701
Authors: Lapin J, Awosanya EO, Esteves RJA, Nevzorov AA
Abstract
The benefits of triple-resonance experiments for structure determination of macroscopically oriented membrane proteins by solid-state NMR are discussed. While double-resonance 1H/15N experiments are effective for structure elucidation of alpha-helical domains, extension of the method of oriented samples to more complex topologies and assessing side-chain conformations necessitates further development of triple-resonance (1H/13C/15N) NMR pulse sequences. Incorporating additional spectroscopic dimensions involving 13C spin-bearing nuclei, however, introduces essential complications arising from the wide frequency range of the 1H-13C dipolar couplings and 13C CSA (>20*kHz), and the presence of the 13C-13C homonuclear dipole-dipole interactions. The recently reported ROULETTE-CAHA pulse sequence, in combination with the selective z-filtering, can be used to evolve the structurally informative 1H-13C dipolar coupling arising from the aliphatic carbons while suppressing the signals from the carbonyl and methyl regions. Proton-mediated magnetization transfer under mismatched Hartman-Hahn conditions (MMHH) can be used to correlate 13C and 15N nuclei in such triple-resonance experiments for the subsequent 15N detection. The recently developed pulse sequences are illustrated for n-acetyl Leucine (NAL) single crystal and doubly labeled Pf1 coat protein reconstituted in magnetically aligned bicelles. An interesting observation is that in the case of 15N-labeled NAL measured at 13C natural abundance, the triple (1H/13C/15N) MMHH scheme predominantly gives rise to long-range intermolecular magnetization transfers from 13C to 15N spins; whereas direct Hartmann-Hahn 13C/15N transfer is entirely intramolecular. The presented developments advance NMR of oriented samples for structure determination of membrane proteins and liquid crystals.
PMID: 33260039 [PubMed - as supplied by publisher]
[NMR paper] A theoretical assessment of structure determination of multi-span membrane proteins by oriented sample solid-state NMR spectroscopy.
A theoretical assessment of structure determination of multi-span membrane proteins by oriented sample solid-state NMR spectroscopy.
Related Articles A theoretical assessment of structure determination of multi-span membrane proteins by oriented sample solid-state NMR spectroscopy.
Aust J Chem. 2020;73(3):246-251
Authors: Weber DK, Veglia G
Abstract
Oriented sample solid state NMR (OS-ssNMR) spectroscopy allows direct determination of the structure and topology of membrane proteins reconstituted into aligned lipid bilayers. While...
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[NMR paper] Macrodiscs Comprising SMALPs for Oriented Sample Solid-State NMR Spectroscopy of Membrane Proteins.
Macrodiscs Comprising SMALPs for Oriented Sample Solid-State NMR Spectroscopy of Membrane Proteins.
Related Articles Macrodiscs Comprising SMALPs for Oriented Sample Solid-State NMR Spectroscopy of Membrane Proteins.
Biophys J. 2018 Jun 15;:
Authors: Radoicic J, Park SH, Opella SJ
Abstract
Macrodiscs, which are magnetically alignable lipid bilayer discs with diameters of >30*nm, were obtained by solubilizing protein-containing liposomes with styrene-maleic acid copolymers. Macrodiscs provide a detergent-free phospholipid bilayer...
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Macrodiscs Comprising SMALPs for Oriented Sample Solid-State NMR Spectroscopy of Membrane Proteins
Macrodiscs Comprising SMALPs for Oriented Sample Solid-State NMR Spectroscopy of Membrane Proteins
Publication date: Available online 15 June 2018
Source:Biophysical Journal</br>
Author(s): Jasmina Radoicic, Sang Ho Park, Stanley J. Opella</br>
Macrodiscs, which are magnetically alignable lipid bilayer discs with diameters of >30*nm, were obtained by solubilizing protein-containing liposomes with styrene-maleic acid copolymers. Macrodiscs provide a detergent-free phospholipid bilayer environment for biophysical and functional studies of membrane proteins under...
[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...
1H/13C/15N triple-resonance experiments for structure determinaton of membrane proteins by oriented-sample NMR.
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