Selective (1)H-(1)H Distance Restraints in Fully Protonated Proteins by Very Fast Magic-Angle Spinning Solid-State NMR.
 

Selective (1)H-(1)H Distance Restraints in Fully Protonated Proteins by Very Fast Magic-Angle Spinning Solid-State NMR.

Related Articles Selective (1)H-(1)H Distance Restraints in Fully Protonated Proteins by Very Fast Magic-Angle Spinning Solid-State NMR.

J Phys Chem Lett. 2017 May 11;:

Authors: Jain MG, Lalli D, Stanek J, GOwda CM, Prakash S, Schwarzer TS, Schubeis T, Castiglione K, Andreas LB, Madhu PK, Pintacuda G, Agarwal V

Abstract
Very fast magic-angle spinning (MAS>80 kHz) NMR spectroscopy combined with high field magnets has enabled the acquisition of proton-detected spectra in fully protonated solid samples with sufficient resolution and sensitivity. One of the primary challenges in structure determination of protein is to observe long-range 1H-1H proximities. We demonstrate a new experiment that allows observing selective HN-HN/H?-H?/Hmethyl-Hmethyl contacts on the order of 5-6 Å despite the presence of other protons at shorter distances. Such contacts are critical to access tertiary structures of proteins but difficult to observe in a dense proton matrix. They have only been reported in perdeuterated proteins. Numerical simulations and experiments show our experiment to outperform the currently used recoupling methods, RFDR, in terms of sensitivity and number of observed long-range contacts. Our method is demonstrated on GB1 and the ?-barrel membrane protein AlkL.


PMID: 28492324 [PubMed - as supplied by publisher]



More...