Morten Bjerring

Unique Identification of Supramolecular Structures in Amyloid Fibrils by Solid-State NMR Spectroscopy†

The fibril structure formed by the amyloidogenic fragment SNNFGAILSS of the human islet amyloid polypeptide (hIAPP) is determined with 0.52 A resolution. Symmetry information contained in the easily obtainable resonance assignments from solid-state NMR spectra (see picture), along with long-range constraints, can be applied to uniquely identify the supramolecular organization of fibrils.

A Hexameric Peptide Barrel as Building Block of Amyloid‐β Protofibrils

Oligomeric and protofibrillar aggregates formed by the amyloid-β peptide (Aβ) are believed to be involved in the pathology of Alzheimers disease. Central to Alzheimer pathology is also the fact that the longer Aβ42 peptide is more prone to aggregation than the more prevalent Aβ40 . Detailed structural studies of Aβ oligomers and protofibrils have been impeded by aggregate heterogeneity and instability. We previously engineered a variant of Aβ that forms stable protofibrils and here we use solid-state NMR spectroscopy and molecular modeling to derive a structural model of these. NMR data are consistent with packing of residues 16 to 42 of Aβ protomers into hexameric barrel-like oligomers within the protofibril. The core of the oligomers consists of all residues of the central and C-terminal hydrophobic regions of Aβ, and hairpin loops extend from the core. The model accounts for why Aβ42 forms oligomers and protofibrils more easily than Aβ40 .

Optimal (2)H rf Pulses and (2)H-(13)C Cross-Polarization Methods for Solid-State (2)H MAS NMR of Perdeuterated Proteins.

We present a novel concept for rf pulses and optimal control designed cross-polarization experiments for quadrupolar nuclei. The methods are demonstrated for (2)H CP-MAS and (2)H multiple-pulse NMR of perdeuterated proteins, for which sensitivity enhancements up to an order of magnitude are presented relative to commonly used approaches. The so-called RESPIRATION rf pulses combines the concept of short broad-band pulses with generation of pulses with large flip angles through distribution of the rf pulse over several rotor echoes. This lead to close-to-ideal rf pulses, facilitating implementation of experiments relying on the ability to realize high-performance 90 and 180° pulses, as, for example, in refocused INEPT and double-to-single quantum coherence experiments, or just pulses that provide a true representation of the quadrupolar powder pattern to extract information about the structure or dynamics. The optimal control (2)H → (13)C CP-MAS method demonstrates transfer efficiencies up to around 85% while being extremely robust toward rf inhomogeneity and resonance offsets.

Fullerene alloy formation and the benefits for efficient printing of ternary blend organic solar cells

Composition average dependent properties for blends of the conjugated polymer P3HT and the fullerenes [60]PCBM, [60]ICBA and their mixtures were studied using cross-polarization magic-angle-spinning solid-state NMR techniques. We found that the blended fullerenes form an alloy and that when mixed with a third polymer component, the system exhibits pseudo-binary phase behaviour instead of the expected ternary phase behaviour. Our results experimentally confirm the earlier hypothesis that the unexpected composition average dependent IV-behaviour for these supposed ternary mixtures are indeed due to them behaving as pseudo-binary mixtures due to alloying of the fullerene components. This finding has vast implications for the understanding of polymer–fullerene mixtures and quite certainly also their application in organic solar cells where performance hinges critically on the blend behaviour which is also investigated in this study.

A Comparison of NCO and NCA Transfer Methods for Biological Solid-State NMR Spectroscopy

Three different techniques (adiabatic passage Hartman-Hahn cross-polarization, optimal control designed pulses, and EXPORT) are compared for transferring (15)N magnetization to (13)C in solid-state NMR experiments under magic-angle-spinning conditions. We demonstrate that, in comparison to adiabatic passage Hartman-Hahn cross-polarization, optimal control transfer pulses achieve similar or better transfer efficiencies for uniformly-(13)C,(15)N labeled samples and are generally superior for samples with non-uniform labeling schemes (such as 1,3- and 2-(13)C glycerol labeling). In addition, the optimal control pulses typically use substantially lower average RF field strengths and are more robust with respect to experimental variation and RF inhomogeneity. Consequently, they are better suited for demanding samples.

Refocused continuous-wave decoupling: A new approach to heteronuclear dipolar decoupling in solid-state NMR spectroscopy

A novel strategy for heteronuclear dipolar decoupling in magic-angle spinning solid-state nuclear magnetic resonance (NMR) spectroscopy is presented, which eliminates residual static high-order terms in the effective Hamiltonian originating from interactions between oscillating dipolar and anisotropic shielding tensors. The method, called refocused continuous-wave (rCW) decoupling, is systematically established by interleaving continuous wave decoupling with appropriately inserted rotor-synchronized high-power π refocusing pulses of alternating phases. The effect of the refocusing pulses in eliminating residual effects from dipolar coupling in heteronuclear spin systems is rationalized by effective Hamiltonian calculations to third order. In some variants the π pulse refocusing is supplemented by insertion of rotor-synchronized π/2 purging pulses to further reduce the residual dipolar coupling effects. Five different rCW decoupling sequences are presented and their performance is compared to state-of-the-art decoupling methods. The rCW decoupling sequences benefit from extreme broadbandedness, tolerance towards rf inhomogeneity, and improved potential for decoupling at relatively low average rf field strengths. In numerical simulations, the rCW schemes clearly reveal superior characteristics relative to the best decoupling schemes presented so far, which we to some extent also are capable of demonstrating experimentally. A major advantage of the rCW decoupling methods is that they are easy to set up and optimize experimentally.

Heteronuclear coherence transfer in solid-state nuclear magnetic resonance using a γ-encoded transferred echo experiment

A novel type of solid-state nuclear magnetic resonance experiment for efficient transfer of coherence between different nuclear spin I=1/2 species under magic-angle spinning conditions is introduced. The method combines the attractive features of γ-encoded dipolar recoupling [Nielsen et al., J. Chem. Phys. 101, 1805 (1995)] with coherence transfer mediated by a longitudinal spin-order operator in a transferred echo experiment. Using two-channel rotary resonance recoupling with different phase and amplitude modulation schemes, the transferred echo sequence can be tuned to achieve dipolar recoupling and coherence transfer over a well-defined range of chemical shifts while keeping the ratio between the rf field strength and the sample spinning frequency relatively low. The method, referred to as gamma-encoded transfer echo, is described analytically, by numerical simulations for various different spin systems, and experimentally by 15N to 13C coherence transfers in a powder sample of 13C, 15N-labeled glycine.

In Situ Solid‐State NMR Spectroscopy of Protein in Heterogeneous Membranes: The Baseplate Antenna Complex of Chlorobaculum tepidum

A clever combination: an in situ solid-state NMR analysis of CsmA proteins in the heterogeneous environment of the photoreceptor of Chlorobaculum tepidum is reported. Using different combinations of 2D and 3D solid-state NMR spectra, 90 % of the CsmA resonances are assigned and provide on the basis of chemical shift data information about the structure and conformation of CsmA in the CsmA-bacteriochlorophyll a complex.

Efficient and Robust Heteronuclear Cross-Polarization for High-Speed-Spinning Biological Solid-State NMR Spectroscopy

We present a new and highly efficient approach for heteronuclear coherence transfer in solid-state NMR spectroscopy under high-speed spinning conditions. The so-called (RESPIRATION)CP experiment exploits phase-alternated recoupling on only one of the two rf channels intertwined in a synchronized train of short rf pulses on both channels. The method provides significantly higher efficiencies than state-of-the art techniques including ramped and adiabatic cross-polarization experiments with long durations of intense rf irradiation. At the same time, it is easier to setup experimentally and significantly more robust toward imperfections such as rf inhomogeneity, misadjustments, and sample-induced variations in the rf tuning. The method is described analytically, numerically, and experimentally for biological solids. We demonstrate sensitivity gains of factors of 1.3 and 1.8 for typical (1)H→(15)N and (15)N→(13)C transfers and a combined gain of a factor of 2-4 for a typical NCA experiment for biological solid-state NMR.

Solid-state NMR heteronuclear dipolar recoupling using off-resonance symmetry-based pulse sequences

Abstract A new approach to heteronuclear dipolar recoupling in solid-state NMR which combines symmetry-based, γ-encoded dipolar recoupling with off-resonance rf irradiation is described. By irradiating the two spin species with offsets of equal magnitude but opposite sign, the dipolar scaling factor may be increased while using lower rf field strengths than under on-resonance conditions. Larger scaling makes the experiment less susceptible to relaxation, while released rf field requirements facilitate combination with fast magic-angle spinning while maintaining efficient 1 H decoupling. Furthermore, homonuclear dipolar couplings may be suppressed by adjusting the off-resonance irradiation to the Lee–Goldburg condition.