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Cell-type-specific labeling and profiling of glycans in living mice

ABSTRACT: Metabolic labeling of glycans with clickable unnatural sugars has enabled glycan imaging and glycoproteomics in mixed cell populations. However, in vivo labeling and profiling of cell-type-specific glycans remains challenging. Here, we develop genetically encoded metabolic glycan labeling (GeMGL), a cell-type-specific strategy based on a bump-hole pair of an unnatural sugar and its matching engineered enzyme. N pentynylacetylglucosamine (GlcNAl) serves as a bumped analog of N acetylglucosamine (GlcNAc) that can be specifically incorporated into glycans of cells expressing an engineered mutant of UDP-GlcNAc pyrophosphorylase, AGX2F383G. GeMGL with the 1,3-di-O-propionylated GlcNAl (1,3-Pr2GlcNAl) and AGX2F383G pair is demonstrated in cell co-cultures, and used for specific labeling of glycans in mouse xenograft tumors. By generating a transgenic mouse line with AGX2F383G expressed under cardiomyocyte-specific promoter, we perform specific imaging of cardiomyocyte glycans in the heart and identified 582 cardiomyocyte O-GlcNAcylated proteins with no interference from other cardiac cell types. GeMGL will facilitate cell-type-specific glycoproteomics in various tissues and disease models.

INSTRUMENT(S): LTQ Orbitrap Elite, Q Exactive

ORGANISM(S): Homo Sapiens (human) Mus Musculus (mouse)

TISSUE(S): Heart, Permanent Cell Line Cell, Cell Culture

DISEASE(S): Breast Cancer

SUBMITTER: Qitao Song

LAB HEAD: Xing Chen

PROVIDER: PXD030663 | Pride | 2022-02-28

REPOSITORIES: Pride

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