Deuterated Peptides and Proteins: Structure and Dynamics Studies by MAS Solid-state NMR

Reif, Bernd

doi:10.1002/9780470034590.emrstm1080

Cited by 4 publications

(3 citation statements)

References 79 publications

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“…MAS NMR has been used for studying dynamics of aromatic rings ( Gall et al, 1981 , Gall et al, 1982 , Vugmeyster et al, 2015 , Vugmeyster et al, 2017 ) and protein dynamics more generally (see reviews, e.g. references ( Bonaccorsi et al, 2021 , Krushelnitsky et al, 2013 , Krushelnitsky and Reichert, 2005 , Lewandowski, 2013 , Reif, 2012 , Schanda and Ernst, 2016 , Watt and Rienstra, 2014 , Yan et al, 2013 )). We have recently applied a selective isotope-labeling strategy combined with sensitive proton-detected MAS NMR pulse sequences to quantitatively probe aromatic ring dynamics over a wide range of time scales ( Gauto et al, 2019 ).…”

Section: Introductionmentioning

confidence: 99%

Aromatic ring flips in differently packed ubiquitin protein crystals from MAS NMR and MD

Gauto¹,

Lebedenko²,

Becker³

et al. 2023

Journal of Structural Biology: X

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Section: Introductionmentioning

confidence: 99%

Aromatic ring flips in differently packed ubiquitin protein crystals from MAS NMR and MD

Gauto¹,

Lebedenko²,

Becker³

et al. 2023

Journal of Structural Biology: X

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“…MAS NMR has been used for studying dynamics of aromatic rings (5,(17)(18)(19) and protein dynamics more generally (see reviews, e.g. references (20)(21)(22)(23)(24)(25)(26)(27)). We have recently applied a selective isotope-labeling strategy combined with sensitive proton-detected MAS NMR pulse sequences to quantitatively probe aromatic ring dynamics over a wide range of time scales (28).…”

Section: Introductionmentioning

confidence: 99%

Aromatic ring flips in differently packed ubiquitin protein crystals from MAS NMR and MD

Gauto

Lebedenko

Becker

et al. 2022

Preprint

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Probing the dynamics of aromatic side chains provides important insights into the behavior of a protein because flips of aromatic rings in a protein's hydrophobic core report on breathing motion involving large part of the protein. Inherently invisible to crystallography, aromatic motions have been primarily studied by solution NMR. Here we apply magic-angle spinning NMR, advanced phenylalanine 1H-13C/2H isotope labeling and molecular simulations to a protein in three different crystal packing environments to shed light onto possible impact of packing on ring flips. We find that ring flips in ubiquitin are quite conserved in the different crystal forms, even though the intermolecular packing is quite different suggesting that intramolecular influences are more important than intermolecular (packing) effects.

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“…In comparison, the large size of the 2 H quadrupolar coupling interaction makes it a natural probe of large-amplitude molecular motions and small order parameters. Moreover, the increasing use of perdeuterated proteins and other biomolecules for 1 H-detected structure determination experiments (Andreas et al 2015; Reif 2012) makes it efficient to characterize molecular motions using the same samples. To obtain 2 H quadrupolar spectra with site resolution, indirect detection of 2 H spectra through 13 C and/or 15 N is necessary, which requires coherence transfer from 2 H to 13 C/ 15 N (Hologne et al 2005).…”

Section: Introductionmentioning

confidence: 99%

2H–13C correlation solid-state NMR for investigating dynamics and water accessibilities of proteins and carbohydrates

et al. 2017

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Site-specific determination of molecular motion and water accessibility by indirect detection of H NMR spectra has advantages over dipolar-coupling based techniques due to the large quadrupolar couplings and the ensuing high angular resolution. Recently, a Rotor Echo Short Pulse IRrAdiaTION mediated cross polarization (CP) technique was developed, which allowed efficient transfer of H magnetization toC at moderate H radiofrequency field strengths available on most commercial MAS probes. In this work, we investigate theH-C magnetization transfer characteristics of one-bond perdeuterated CD spin systems and two-bond H/D exchanged C-(O)-D and C-(N)-D spin systems in carbohydrates and proteins. Our results show that multi-bond, broadbandH-C polarization transfer can be achieved using H radiofrequency fields of ~50 kHz, relatively short contact times of 1.3-1.7 ms, and with sufficiently high sensitivity to enable 2DH-C correlation experiments with undistorted H spectra in the indirect dimension. To demonstrate the utility of thisH-C technique for studying molecular motion, we show H-C correlation spectra of perdeuterated bacterial cellulose, whose surface glucan chains exhibit a motionally averaged C6 H quadrupolar coupling that indicates fast trans-gauche isomerization about the C5-C6 bond. In comparison, the interior chains in the microfibril core are fully immobilized. Application of theH-C correlation experiment to H/D exchanged Arabidopsis primary cell walls show that the O-D quadrupolar spectra of the highest polysaccharide peaks can be fit to a two-component model, in which 74% of the spectral intensity, assigned to cellulose, has a near-rigid-limit coupling, while 26% of the intensity, assigned to matrix polysaccharides, has a weakened coupling of 50 kHz. The latter O-D quadrupolar order parameter of 0.22 is significantly smaller than previously reported C-D dipolar order parameters of 0.46-0.55 for pectins, suggesting that additional motions exist at the C-O bonds in the wall polysaccharides. H-C polarization transfer profiles are also compared between statistically deuterated and H/D exchanged GB1.

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Deuterated Peptides and Proteins: Structure and Dynamics Studies by MAS Solid-state NMR

Cited by 4 publications

References 79 publications

Aromatic ring flips in differently packed ubiquitin protein crystals from MAS NMR and MD

Aromatic ring flips in differently packed ubiquitin protein crystals from MAS NMR and MD

Aromatic ring flips in differently packed ubiquitin protein crystals from MAS NMR and MD

2H–13C correlation solid-state NMR for investigating dynamics and water accessibilities of proteins and carbohydrates

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