Exploring amyloid formation by a de novo design.

Protein deposition as amyloid fibrils underlies many debilitating human disorders. The complexity and size of disease-related polypeptides, however, often hinders a detailed rational approach to study effects that contribute to the process of amyloid formation. We report here a simplified peptide sequence successfully designed de novo to fold into a coiled-coil conformation under ambient conditions but to transform into amyloid fibrils at elevated temperatures.

NMR spectra of a microcrystalline protein at 30 kHz MAS.

Proteins are not always available in amounts desirable for solid-state magic-angle spinning (MAS) nuclear-magnetic resonance (NMR) spectroscopy. To maximize the signal-to-noise ratio achievable with small samples, the filling factor must be optimized by using small-diameter MAS rotors. These rotors have the added benefit of allowing higher radio frequency field amplitudes during polarization transfer steps and during decoupling periods as well as allowing higher spinning frequencies.

Fast-MAS total through-bond correlation spectroscopy using adiabatic pulses.

Pulse sequences consisting of adiabatic pulses for total through-bond correlation spectroscopy (TOBSY) under magic-angle spinning (MAS) are introduced. Above a certain threshold, the polarization transfer achieved with these sequences is largely insensitive to the amplitude and homogeneity of the radiofrequency field employed. An experimental transfer efficiency of up to 76% was achieved in a two-spin system using the sequence WiW9(24)(1) at a MAS frequency of 26.