Display of the Human Mucinome with Defined O-Glycans by Gene Engineered Cells

ABSTRACT: 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 can form aggregated networks and bundles that serve as a barrier, but also assist in containing, feeding, clearing and replenishing microorganisms. 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. However, the O-glycodomains have long evaded detailed structural and functional exploration. Here, we developed a cell-based platform for the display and production of human TR O-glycodomains (~200 amino acids) with tunable structures and patterns of O-glycans using membrane-bound and secreted constructs expressed in glycoengineered HEK293 cells. The expressed mucin TRs revealed surprisingly high fidelity in complete decoration with designed O-glycan structures, which enabled intact mass analysis with the simplest glycoforms. 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.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Sergey Vakhrushev  

LAB HEAD: Sergey Vakhrushev

PROVIDER: PXD024851 | Pride | 2021-05-11

REPOSITORIES: Pride