Mixed pyruvate labeling enables backbone resonance assignment of large proteins using a single experiment.

ABSTRACT: Backbone resonance assignment is a critical first step in the investigation of proteins by NMR. This is traditionally achieved with a standard set of experiments, most of which are not optimal for large proteins. Of these, HNCA is the most sensitive experiment that provides sequential correlations. However, this experiment suffers from chemical shift degeneracy problems during the assignment procedure. We present a strategy that increases the effective resolution of HNCA and enables near-complete resonance assignment using this single HNCA experiment. We utilize a combination of 2-¹³C and 3-¹³C pyruvate as the carbon source for isotope labeling, which suppresses the one bond (¹J_αβ) coupling providing enhanced resolution for the Cα resonance and a

SUBMITTER: Robson SA 

PROVIDER: S-EPMC5783931 | biostudies-literature | 2018 Jan

REPOSITORIES: biostudies-literature