Sequential Glycosylation of Proteins with Substrate-Specific N-Glycosyltransferases.

ABSTRACT: Protein glycosylation is a common post-translational modification that influences the functions and properties of proteins. Despite advances in methods to produce defined glycoproteins by chemoenzymatic elaboration of monosaccharides, the understanding and engineering of glycoproteins remain challenging, in part, due to the difficulty of site-specifically controlling glycosylation at each of several positions within a protein. Here, we address this limitation by discovering and exploiting the unique, conditionally orthogonal peptide acceptor specificities of N-glycosyltransferases (NGTs). We used cell-free protein synthesis and mass spectrometry of self-assembled monolayers to rapidly screen 41 putative NGTs and rigorously characterize the unique acceptor sequence preferences of fou

SUBMITTER: Lin L 

PROVIDER: S-EPMC7047269 | biostudies-literature | 2020 Feb

REPOSITORIES: biostudies-literature