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

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 an acid cleavable biotinylated tag. Glycoproteins are enriched using Streptavidin and. after on-bead digestion, glycopeptides are eluted in acid-conditions to be then analysed by mass spectrometry.

INSTRUMENT(S): Orbitrap Eclipse

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

SUBMITTER: Benjamin Schumann

LAB HEAD: Benjamin Schumann

PROVIDER: PXD035445 | Pride | 2022-09-10

REPOSITORIES: Pride

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			Action	DRS
	IQ2366_SampleList.csv	Csv
	IQ2366_SampleList.numbers	Other
	IQ2366_ciocea_CO-CULTURE_G1_MCF7-mock_4T1-mock_DMSO.raw	Raw
	IQ2366_ciocea_CO-CULTURE_G2_MCF7-mock_4T1-BOCTAG_Ac4GalNAlk6-10uM.raw	Raw
	IQ2366_ciocea_CO-CULTURE_G3_MCF7-BOCTAG_4T1-BOCTAG_DMSO.raw	Raw

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