Reshaping of a Glycoside Hydrolase Active Site through Expression-Compensated Droplet-Based Microfluidic Screening Provides Useful Tools for Glycomics.

ABSTRACT: The glycosylation of proteins endows them with distinct biophysical properties and allows them to play fundamental roles in cellular communication. Much of our understanding of glycoproteins has derived from the ability to enzymatically manipulate glycan structures. In particular, selective cleavage of glycans from proteins simplifies the analysis of glycoproteins and the determination of structure-activity relationships. However, limited enzymatic tools are available for the study of mucin-type O-glycans. To address this, we carried out the directed evolution of a glycoside hydrolase to increase its ability to cleave the sialyl T-antigen, a ubiquitous O-glycan structure in humans. We employed ultrahigh-throughput droplet-based microfluidics to rapidly screen vast libraries of variants in

SUBMITTER: Wardman JF 

PROVIDER: S-EPMC12550630 | biostudies-literature | 2025 Oct

REPOSITORIES: biostudies-literature