Display of the human mucinome with defined O-glycans by gene engineered cells

Mucins are a large family of heavily O-glycosylated proteins that cover all mucosal surfaces and constitute the major macromolecules in most body fluids. Mucins are primarily defined by their variable tandem repeat (TR) domains that are densely decorated with different O-glycan structures in distinct patterns, and these arguably convey much of the informational content of mucins. Here, we develop a cell-based platform for the display and production of human TR O-glycodomains (similar to 200 amino acids) with tunable structures and patterns of O-glycans using membrane-bound and secreted reporters expressed in glycoengineered HEK293 cells. Availability of defined mucin TR O-glycodomains advances experimental studies into the versatile role of mucins at the interface with pathogenic microorganisms and the microbiome, and sparks new strategies for molecular dissection of specific roles of adhesins, glycoside hydrolases, glycopeptidases, viruses and other interactions with mucin TRs as highlighted by examples.

Automated summary

Mucins are heavily O-glycosylated proteins that are found on mucosal surfaces and in body fluids. A cell-based platform has been developed to display and produce human TR O-glycodomains with tunable structures and patterns of O-glycans, allowing for experimental studies on the versatile role of mucins in interactions with pathogenic microorganisms and the microbiome.