Selective isotope labeling for NMR structure determination of proteins in complex with unlabeled ligands.

ABSTRACT: The physiological role of proteins is frequently linked to interactions with non-protein ligands or posttranslational modifications. Structural characterization of these complexes or modified proteins by NMR may be difficult as the ligands are usually not available in an isotope-labeled form and NMR spectra may suffer from signal overlap. Here, we present an optimized approach that uses specific NMR isotope-labeling schemes for overcoming both hurdles. This approach enabled the high-resolution structure determination of the farnesylated C-terminal domain of the peroxisomal protein PEX19. The approach combines specific ¹³C, ¹⁵N and ²H isotope labeling with tailored NMR experiments to (i) unambiguously identify the NMR frequencies and the stereochemistry of t

SUBMITTER: Tripsianes K 

PROVIDER: S-EPMC6525670 | biostudies-literature | 2019 Apr

REPOSITORIES: biostudies-literature