MAS NMR detection of hydrogen bonds for protein secondary structure characterization

Stöppler, Daniel; Perodeau, Jacqueline; Nieuwkoop, Andrew J.; Oschkinat, Hartmut

doi:10.1007/s10858-020-00307-z

Cited by 14 publications

(15 citation statements)

References 42 publications

(42 reference statements)

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“…Similar observations are e.g. observed in (Friedrich et al 2020 ). The spin diffusion under spin-lock condition during the CP actually seems to provide efficient polarization transfer, despite the scaling of the homonuclear dipolar couplings by a factor of −0.5 (Rhim et al 1970 ).…”

Section: Resultssupporting

confidence: 92%

Biomolecular solid-state NMR spectroscopy at 1200 MHz: the gain in resolution

et al. 2021

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Progress in NMR in general and in biomolecular applications in particular is driven by increasing magnetic-field strengths leading to improved resolution and sensitivity of the NMR spectra. Recently, persistent superconducting magnets at a magnetic field strength (magnetic induction) of 28.2 T corresponding to 1200 MHz proton resonance frequency became commercially available. We present here a collection of high-field NMR spectra of a variety of proteins, including molecular machines, membrane proteins, viral capsids, fibrils and large molecular assemblies. We show this large panel in order to provide an overview over a range of representative systems under study, rather than a single best performing model system. We discuss both carbon-13 and proton-detected experiments, and show that in 13C spectra substantially higher numbers of peaks can be resolved compared to 850 MHz while for 1H spectra the most impressive increase in resolution is observed for aliphatic side-chain resonances.

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Section: Resultssupporting

confidence: 92%

Biomolecular solid-state NMR spectroscopy at 1200 MHz: the gain in resolution

et al. 2021

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“…A 9 ms 13 CO-1 H CP-step was employed to achieve a polarization transfer to both NH 2 protons of the neighboring asparagine sidechain within the same ladder. Similar experiments have been described recently to identify backbone hydrogen bonding patterns in SH3 and GB1 (Friedrich et al, 2020). And indeed, the 15 N planes at the NH 2 side-chain chemical-shift value of N262 (112.7 ppm) and N226 (114.8 ppm) clearly show cross peaks to the proton shifts of the NH 2 group of N226 and N262, respectively (see Figure 2A, red arrows) indicating their spatial proximity.…”

Section: Asparagine Ladderssupporting

confidence: 85%

Asparagine and Glutamine Side-Chains and Ladders in HET-s(218–289) Amyloid Fibrils Studied by Fast Magic-Angle Spinning NMR

Wiegand

Malär

Cadalbert

et al. 2020

Front. Mol. Biosci.

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Asparagine and glutamine side-chains can form hydrogen-bonded ladders which contribute significantly to the stability of amyloid fibrils. We show, using the example of HET-s(218-289) fibrils, that the primary amide side-chain proton resonances can be detected in cross-polarization based solid-state NMR spectra at fast magic-angle spinning (MAS). J-coupling based experiments offer the possibility to distinguish them from backbone amide groups if the spin-echo lifetimes are long enough, which turned out to be the case for the glutamine side-chains, but not for the asparagine sidechains forming asparagine ladders. We explore the sensitivity of NMR observables to asparagine ladder formation. One of the two possible asparagine ladders in HETs(218-289), the one comprising N226 and N262, is assigned by proton-detected 3D experiments at fast MAS and significant de-shielding of one of the NH 2 proton resonances indicative of hydrogen-bond formation is observed. Small rotating-frame 15 N relaxation-rate constants point to rigidified asparagine side-chains in this ladder. The proton resonances are homogeneously broadened which could indicate chemical exchange, but is presently not fully understood. The second asparagine ladder (N243 and N279) in contrast remains more flexible.

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“…Similarly, collective exchange processes in the active-site proton cage in bacteriorhodopsin could be characterized . Furthermore, backbone hydrogen bonds that stabilize protein secondary structure elements in solid proteins could be identified . The same experiment applied to side chain resonances showed that salt bridges stabilize amyloid quarternary structure .…”

Section: Narrow Proton Resonances By Spin Dilutionmentioning

confidence: 98%

Deuteration for High-Resolution Detection of Protons in Protein Magic Angle Spinning (MAS) Solid-State NMR

Reif

2021

Chem. Rev.

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Proton detection developed in the last 20 years as the method of choice to study biomolecules in the solid state. In perdeuterated proteins, proton dipolar interactions are strongly attenuated, which allows yielding of high-resolution proton spectra. Perdeuteration and backsubstitution of exchangeable protons is essential if samples are rotated with MAS rotation frequencies below 60 kHz. Protonated samples can be investigated directly without spin dilution using proton detection methods in case the MAS frequency exceeds 110 kHz. This review summarizes labeling strategies and the spectroscopic methods to perform experiments that yield assignments, quantitative information on structure, and dynamics using perdeuterated samples. Techniques for solvent suppression, H/D exchange, and deuterium spectroscopy are discussed. Finally, experimental and theoretical results that allow estimation of the sensitivity of proton detected experiments as a function of the MAS frequency and the external B 0 field in a perdeuterated environment are compiled.

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MAS NMR detection of hydrogen bonds for protein secondary structure characterization

Cited by 14 publications

References 42 publications

Biomolecular solid-state NMR spectroscopy at 1200 MHz: the gain in resolution

Biomolecular solid-state NMR spectroscopy at 1200 MHz: the gain in resolution

Asparagine and Glutamine Side-Chains and Ladders in HET-s(218–289) Amyloid Fibrils Studied by Fast Magic-Angle Spinning NMR

Deuteration for High-Resolution Detection of Protons in Protein Magic Angle Spinning (MAS) Solid-State NMR

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