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

Lars Hansen, Yoshiki Narimatsu, Henrik Clausen, Sergey Y Vakhrushev, Bernard Henrissat, Renaud Vincentelli, Cornelis A M de Haan, Erik de Vries, Ajit Varki, Paul M Sullam, Barbara A Bensing, Tina M Iverson, Leonor David, Rebecca Nason, Leo Alexander Dworkin, Hiren J Joshi, Ulla Mandel, Sanae Furukawa, Fabien Durbesson, Daniel M Sørensen, Wenjuan Du, Adnan Halim, Zilu Ye, Lingbo Sun, Andriana Konstantinidi, Christian Büll

Journal: Nature communications 2021;12(1):4070

PMID: 34210959

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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 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 (~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.

Address: Copenhagen Center for Glycomics, Departments of Cellular and Molecular Medicine and Odontology, Faculty of Health Sciences, University of Copenhagen, Copenhagen, Denmark.; Section Virology, Division of Infectious Diseases and Immunology, Department Biomolecular Health Sciences, Faculty of Veterinary Medicine, Utrecht University, CL, Utrecht, the Netherlands.; Architecture et Fonction des Macromolécules Biologiques, CNRS, Aix-Marseille Université, Marseille, France.; Institute of Molecular Pathology and Immunology of the University of Porto/I3S, Porto, Portugal.; Medical Faculty of the University of Porto, Porto, Portugal.; Departments of Pharmacology and Biochemistry, Vanderbilt University, Nashville, TN, USA.; Department of Medicine, The San Francisco Veterans Affairs Medical Center, and the University of California, San Francisco, CA, USA.; The Glycobiology Research and Training Center, and the Department of Cellular and Molecular Medicine, University of California, San Diego, CA, USA.; Department of Biological Sciences, King Abdulaziz University, Jeddah, Saudi Arabia.; Copenhagen Center for Glycomics, Departments of Cellular and Molecular Medicine and Odontology, Faculty of Health Sciences, University of Copenhagen, Copenhagen, Denmark. [email protected].; Copenhagen Center for Glycomics, Departments of Cellular and Molecular Medicine and Odontology, Faculty of Health Sciences, University of Copenhagen, Copenhagen, Denmark. [email protected].; GlycoDisplay ApS, Copenhagen, Denmark. [email protected].

DOI: 10.1038/s41467-021-24366-4