NMR Assignment of Methyl Groups in Immobilized Proteins Using Multiple-Bond ¹³C Homonuclear Transfers, Proton Detection, and Very Fast MAS.

ABSTRACT: In nuclear magnetic resonance spectroscopy of proteins, methyl protons play a particular role as extremely sensitive reporters on dynamics, allosteric effects, and protein-protein interactions, accessible even in high-molecular-weight systems approaching 1 MDa. The notorious issue of their chemical shift assignment is addressed here by a joint use of solid-state ¹H-detected methods at very fast (nearly 100 kHz) magic-angle spinning, partial deuteration, and high-magnetic fields. The suitability of a series of RF schemes is evaluated for the efficient coherence transfer across entire ¹³C side chains of methyl-containing residues, which is key for establishing connection between methyl and backbone ¹H resonances. The performance of ten methods for recoupling

SUBMITTER: Paluch P 

PROVIDER: S-EPMC9002630 | biostudies-literature | 2022

REPOSITORIES: biostudies-literature