Abstract
Proton-detected solid-state NMR (SSNMR) spectroscopy has attracted much attention due to its excellent sensitivity and effectiveness in the analysis of trace amounts of amyloid proteins and other important biological systems. In this perspective article, we present the recent sensitivity limit of (1)H-detected SSNMR using "ultra-fast" magic-angle spinning (MAS) at a spinning rate (ν(R)) of 80-100 kHz. It was demonstrated that the high sensitivity of (1)H-detected SSNMR at ν(R) of 100 kHz and fast recycling using the paramagnetic-assisted condensed data collection (PACC) approach permitted "super-fast" collection of (1)H-detected 2D protein SSNMR. A (1)H-detected 2D (1)H-(15)N correlation SSNMR spectrum for ∼27 nmol of a uniformly (13)C- and (15)N-labeled GB1 protein sample in microcrystalline form was acquired in only 9 s with 50% non-uniform sampling and short recycle delays of 100 ms. Additional data suggests that it is now feasible to detect as little as 1 nmol of the protein in 5.9 h by (1)H-detected 2D (1)H-(15)N SSNMR at a nominal signal-to-noise ratio of five. The demonstrated sensitivity is comparable to that of modern solution protein NMR. Moreover, this article summarizes the influence of ultra-fast MAS and (1)H-detection on the spectral resolution and sensitivity of protein SSNMR. Recent progress in signal assignment and structural elucidation by (1)H-detected protein SSNMR is outlined with both theoretical and experimental aspects.