High and fast: NMR protein–proton side-chain assignments at 160 kHz and 1.2 GHz

Abstract

The NMR resonances of side-chain protons in proteins provide important information, not only about their structure and dynamics, but also about the mechanisms that regulate interactions between macromolecules. However, in...

Microsecond Timescale Conformational Dynamics of a Small‐Molecule Ligand within the Active Site of a Protein

“…macological binders, for supramolecular chemistry and structural biology of protein-ligand complexes but equally concerns natural or biotechnological processes (inhibitors, substrates, or products), as well as studies involving nucleotides or small DNA or RNA molecules, which pose similar limitations to isotope labelling. We expect that in particular for solid samples, with new prospects for NMR-restrained MD simulations, [25] higher magnetic fields, and increasing MAS rates, [26] an exploitation of 1 H relaxation dispersion will bring a greenfield of hitherto inaccessible applications.…”

Konformationeller Austausch eines Liganden in der Bindetasche eines Proteins auf der Mikrosekundenzeitskala

“…3E zeigt drei repräsentative Zeitkorrelationsfunktionen (Time Correlation Function, TCF) für verschiedene (intra-und inter-Ring-)Proton-Proton-Vektoren, die aus den MD-Simulationen wie von Hoffmann et al [24] beschrieben erhalten wurden. [25] stärkere Magnetfelder und zunehmende MAS-Frequenzen [26] Die chemischen Verschiebungen und die Röntgenkristallstrukturdaten sind in den Datenbanken der BMRB (ID 52196) und der PDB (ID 8R1I) zu finden.…”

Using Abundant 1H Polarization to Enhance the Sensitivity of Solid-State NMR Spectroscopy

“…In the meantime, thanks to the tremendous advances in probe technology, faster MAS ,, has enabled the achievement of narrower 1 H peaks and thus higher resolution and sensitivity of 1 H spectra. As a result, proton detection in solid-state NMR has gained dramatic attention in recent years . Specifically, the latest probe technology has enabled the spinning frequency over 180 kHz using small rotors of diameters less than 0.4 mm, ,− under which condition the proton couplings have been suppressed to an impressive level, allowing detailed structural and dynamics investigation of molecular systems with increasing complexity.…”

“…As a result, proton detection in solid-state NMR has gained dramatic attention in recent years . Specifically, the latest probe technology has enabled the spinning frequency over 180 kHz using small rotors of diameters less than 0.4 mm, ,− under which condition the proton couplings have been suppressed to an impressive level, allowing detailed structural and dynamics investigation of molecular systems with increasing complexity. Due to its higher sensitivity, proton detection, exploiting the high natural abundance and gyromagnetic ratio of protons, has been widely incorporated into a broad variety of multidimensional correlation experiments for structural elucidation over the last two decades, especially on proteins by the combined use of paramagnetic relaxation enhancement, ultrahigh magnetic field, , and extensive deuteration. , The higher sensitivity afforded by proton detection has also rendered it possible to observe other insensitive low-γ nuclei signals at fast MAS, such as 14 N, , 15 N, , 35 Cl, 195 Pt, and some other very low gyromagnetic ratio nuclei, even though only a few milligrams of samples (typically around 2 mg for 1.0 mm rotor) are allowed to pack into the rotors. ,− In all proton-detected experimental categories, single-channel 1 H multidimensional NMR spectroscopy is most promising, since it represents one of the most straightforward ways for fully exploiting the high sensitivity of protons to extract structural information .…”

“…As a result, proton detection in solid-state NMR has gained dramatic attention in recent years. 31 latest probe technology has enabled the spinning frequency over 180 kHz using small rotors of diameters less than 0.4 mm, 23,31−33 under which condition the proton couplings have been suppressed to an impressive level, allowing detailed structural and dynamics investigation of molecular systems with increasing complexity. Due to its higher sensitivity, proton detection, exploiting the high natural abundance and gyromagnetic ratio of protons, has been widely incorporated into a broad variety of multidimensional correlation experiments for structural elucidation over the last two decades, 24 especially on proteins by the combined use of paramagnetic relaxation enhancement, 34 ultrahigh magnetic field, 35,36 and extensive deuteration.…”