Accessing isotopically labeled proteins containing genetically encoded phosphoserine for NMR with optimized expression conditions.

ABSTRACT: Phosphoserine (pSer) sites are primarily located within disordered protein regions, making it difficult to experimentally ascertain their effects on protein structure and function. Therefore, the production of ¹⁵N- (and ¹³C)-labeled proteins with site-specifically encoded pSer for NMR studies is essential to uncover molecular mechanisms of protein regulation by phosphorylation. While genetic code expansion technologies for the translational installation of pSer in Escherichia coli are well established and offer a powerful strategy to produce site-specifically phosphorylated proteins, methodologies to adapt them to minimal or isotope-enriched media have not been described. This shortcoming exists because pSer genetic code expansion expression hosts require the genomic

SUBMITTER: Vesely CH 

PROVIDER: S-EPMC9678770 | biostudies-literature | 2022 Oct

REPOSITORIES: biostudies-literature