Proton-detected scalar coupling based assignment strategies in MAS solid-state NMR spectroscopy applied to perdeuterated proteins

Linser, Rasmus; Fink, Uwe; Reif, Bernd

doi:10.1016/j.jmr.2008.04.021

Cited by 101 publications

(111 citation statements)

References 46 publications

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“…[1] On-and off-resonance 13 C pulses were applied as soft rectangular and G3 shaped pulses, respectively. [4,5] Supporting Figure 1. Proton detected NMR experiments.…”

Section: Nmr-spectroscopymentioning

confidence: 99%

Proton‐Detected Solid‐State NMR Spectroscopy of Fibrillar and Membrane Proteins

et al. 2011

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“…[1] On-and off-resonance 13 C pulses were applied as soft rectangular and G3 shaped pulses, respectively. [4,5] Supporting Figure 1. Proton detected NMR experiments.…”

Section: Nmr-spectroscopymentioning

confidence: 99%

Proton‐Detected Solid‐State NMR Spectroscopy of Fibrillar and Membrane Proteins

et al. 2011

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“…We have shown recently that perdeuteration of proteins and recrystallization in a buffer containing varying amounts of D 2 O results in high-resolution MAS solid-state NMR spectra, [27][28][29] which can be employed for solution-state-like correlation experiments such as 3D-HNCACB. 30 Similarly, methyl protons can be employed for spectroscopy as commercially available 2 H-glucose yields an incorporation of -CHD 2 moieties that is on the order of 10%. 31 The sensitivity of the experiments can be increased with precursors such as pyruvate 32 or R-ketoisovalerate.…”

Section: Introductionmentioning

confidence: 99%

Identification of Hydroxyl Protons, Determination of Their Exchange Dynamics, and Characterization of Hydrogen Bonding in a Microcrystallin Protein

et al. 2010

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Abstract:Heteronuclear correlation experiments employing perdeuterated proteins enable the observation of all hydroxyl protons in a microcrystalline protein by MAS solid-state NMR. Dipolar-based sequences allow magnetization transfers that are >50 times faster compared to scalar-coupling-based sequences, which significantly facilitates their assignment. Hydroxyl exchange rates were measured using EXSY-type experiments. We find a biexponential decay behavior for those hydroxyl groups that are involved in side chain-side chain C-O-H · · · OdC hydrogen bonds. The quantification of the distances between the hydroxyl proton and the carbon atoms in the hydrogen-bonding donor as well as acceptor group is achieved via a REDOR experiment. In combination with X-ray data and isotropic proton chemical shifts, availability of 1 H, 13 C distance information can aid in the quantitative description of the geometry of these hydrogen bonds. Similarly, correlations between backbone amide proton and carbonyl atoms are observed, which will be useful in the analysis of the registry of -strand arrangement in amyloid fibrils.

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“…25 At the same time, scalar coupling based experiments yield reliable resonance assignments in the solid-state. [26][27][28][29] Fibril samples prepared from deuterated Ab peptides allow us to collect long range distance restraints which will help to refine the quarternary structure of the fibril. 30,31 In the long run, this labeling scheme will allow an artifact-free quantification of dynamics in the solid-state without spin diffusion as a potential source of error.…”

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Hydrogen bonding involving side chain exchangeable groups stabilizes amyloid quarternary structure

Agarwal

Linser

Dasari

et al. 2013

Phys. Chem. Chem. Phys.

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The amyloid b-peptide (Ab) is the major structural component of amyloid fibrils in the plaques of brains of Alzheimer's disease patients. Numerous studies have addressed important aspects of secondary and tertiary structure of fibrils. In electron microscopic images, fibrils often bundle together. The mechanisms which drive the association of protofilaments into bundles of fibrils are not known. We show here that amino acid side chain exchangeable groups like e.g. histidines can provide useful restraints to determine the quarternary assembly of an amyloid fibril. Exchangeable protons are only observable if a side chain hydrogen bond is formed and the respective protons are protected from exchange. The method relies on deuteration of the Ab peptide. Exchangeable deuterons are substituted with protons, before fibril formation is initiated.Alzheimer's disease (AD) is a slowly progressive neurological disorder which is associated with memory loss, cognitive impairment and finally dementia and death. The amyloid b-peptide (Ab) is the major structural component of amyloid fibrils in the plaques of brains of Alzheimer's disease patients. Ab is a cleavage product resulting from Alzheimer Precursor Protein (APP) processing. 1 The g-secretase cut yields Ab fragments of different lengths of which Ab40 and Ab42 are most abundant. 2,3 In vitro, the Ab peptide aggregates over time into oligomers and fibrillar structures. 4 In the past, several Ab fibril structure models have been suggested. These are based on MAS solidstate NMR experiments, 5-11 and cryo-electron microscopic image reconstructions. 12 Even though there is some controversy concerning the conformational space that Ab fibrils can adopt, all published models, including the 2-fold 7,10 and 3-fold 9 symmetric Ab 1-40 fibril structure suggested by Tycko, and Bertini and co-workers, agree on the basic building block which involves a b-sheet (b1, residues 12-24), a turn, and a second b-sheet (b2, residues 28-40). Biophysical studies indicate a certain degree of conformational plasticity for the N-terminal b-sheet. 13,14 Whereas b2 is present in all the studies, b1 can be truncated or does not adopt a regular b-sheet structure. Depending on the preparation conditions, amide protons in the region of the peptide where the first b-strand is expected show differential H/D protection factors. 13 Similarly, a certain degree of conformational variability in the N-terminus is suggested using cryo-electron microscopy. 14 In all structural models, Ab peptides are arranged in a parallel fashion. An antiparallel arrangement is only observed for the Iowa mutant Ab-D23N, 15 and for truncated forms of the peptide such as Ab [11][12][13][14][15][16][17][18][19][20][21][22][23][24][25] . 16 The fibril structure is stabilized in particular by hydrogen bonding interactions. Therefore, observation of exchangeable and titratable groups is of particular interest to identify hydrogen bonding patterns. We and others could show in the past that high resolution proton spectra can be obtained for...

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Proton-detected scalar coupling based assignment strategies in MAS solid-state NMR spectroscopy applied to perdeuterated proteins

Cited by 101 publications

References 46 publications

Proton‐Detected Solid‐State NMR Spectroscopy of Fibrillar and Membrane Proteins

Proton‐Detected Solid‐State NMR Spectroscopy of Fibrillar and Membrane Proteins

Identification of Hydroxyl Protons, Determination of Their Exchange Dynamics, and Characterization of Hydrogen Bonding in a Microcrystallin Protein

Hydrogen bonding involving side chain exchangeable groups stabilizes amyloid quarternary structure

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