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.
Nat Commun. 2021 Jul 1;12(1):4070. doi: 10.1038/s41467-021-24366-4.
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.
黏蛋白是一类富含 O-糖基化的蛋白大家族，覆盖所有黏膜表面，是大多数体液中的主要大分子。黏蛋白主要通过其可变串联重复（TR）结构域来定义，这些结构域高度装饰有不同的 O-聚糖结构，呈现出独特的模式，这些结构域可以传递黏蛋白的大部分信息内容。在这里，我们开发了一种基于细胞的平台，用于使用糖基工程化的 HEK293 细胞中表达的膜结合和分泌报告蛋白，展示和生产具有可调节结构和 O-聚糖模式的人类 TR O-糖肽结构域（~200 个氨基酸）。具有定义明确的黏蛋白 TR O-糖肽结构域，可促进在与致病性微生物和微生物组的界面处黏蛋白的多功能作用的实验研究，并为分子剖析黏蛋白 TR 与黏附素、糖苷水解酶、糖肽酶、病毒和其他相互作用的特定作用提供新策略，这些作用如示例所示。
