Encoding human serine phosphopeptides in bacteria for proteome-wide identification of phosphorylation-dependent interactions.

ABSTRACT: Post-translational phosphorylation is essential to human cellular processes, but the transient, heterogeneous nature of this modification complicates its study in native systems. We developed an approach to interrogate phosphorylation and its role in protein-protein interactions on a proteome-wide scale. We genetically encoded phosphoserine in recoded E. coli and generated a peptide-based heterologous representation of the human serine phosphoproteome. We designed a single-plasmid library encoding >100,000 human phosphopeptides and confirmed the site-specific incorporation of phosphoserine in >36,000 of these peptides. We then integrated our phosphopeptide library into an approach known as Hi-P to enable proteome-level screens for serine-phosphorylation-dependent human protein interactions

SUBMITTER: Barber KW 

PROVIDER: S-EPMC6590076 | biostudies-literature | 2018 Aug

REPOSITORIES: biostudies-literature