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Cell-type specific tagging of glycoproteins in murine-human co-culture samples

ABSTRACT: We report a strategy termed Bio-Orthogonal Cell line-specific Tagging of Glycoproteins (BOCTAG). Cells are equipped by transfection with an artificial biosynthetic pathway (mut-AGX1/NahK/Nh-GalNAc-T2) that transforms an alkyne tagged sugar into the corresponding nucleotide-sugar. Only transfected cells incorporate the bioorthogonal sugar into glycoproteins in the presence of non-transfected cells. The incorporation of alkyne-sugar in glycoproteins allows the installation, by copper(I)-catalyzed reaction, of a biotinylated tag. Glycoproteins are enriched using Streptavidin, and on-bead digestion yielded a peptide fraction that is analysed by mass spectrometry.

INSTRUMENT(S): Orbitrap Eclipse

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Benjamin Schumann

LAB HEAD: Benjamin Schumann

PROVIDER: PXD035438 | Pride | 2022-09-10

REPOSITORIES: Pride

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Cell-type specific tagging of glycoproteins in murine-human co-culture samples glycoproteomics lysates

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