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K-562 GalNAc-T1/2 Bump-and-hole Whole cell lysate proteomics data

ABSTRACT: Engineered GalNAc-T glycosyltransferases were used to incorporate a chemically modified GalNAc analog into the glycoproteins on the cell surface of K-562 cells. The chemical modification included a bioorthogonal alkyne tag that allowed for introduction of a clickable, acid-cleavable biotin-picolyl-azide. Glycoproteins were enriched using Streptavidin, and on-bead digestion yielded a peptide fraction that was analysed by mass spectrometry.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Suspension Culture

SUBMITTER: Benjamin Schumann

LAB HEAD: Benjamin Schumann

PROVIDER: PXD017989 | Pride | 2020-04-22

REPOSITORIES: Pride

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			Action	DRS
	K562BH-T1DMSO.xlsx	Xlsx
	K562BH-T1_DMSO.raw	Raw
	K562BH-T1cmpd5.raw	Raw
	K562BH-T1cmpd5.xlsx	Xlsx
	K562BH-T2DMSO.raw	Raw

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Publications

Bump-and-Hole Engineering Identifies Specific Substrates of Glycosyltransferases in Living Cells.

Schumann Benjamin B Malaker Stacy Alyse SA Wisnovsky Simon Peter SP Debets Marjoke Froukje MF Agbay Anthony John AJ Fernandez Daniel D Wagner Lauren Jan Sarbo LJS Lin Liang L Li Zhen Z Choi Junwon J Fox Douglas Michael DM Peh Jessie J Gray Melissa Anne MA Pedram Kayvon K Kohler Jennifer Jean JJ Mrksich Milan M Bertozzi Carolyn Ruth CR

Molecular cell 20200422 5

Studying posttranslational modifications classically relies on experimental strategies that oversimplify the complex biosynthetic machineries of living cells. Protein glycosylation contributes to essential biological processes, but correlating glycan structure, underlying protein, and disease-relevant biosynthetic regulation is currently elusive. Here, we engineer living cells to tag glycans with editable chemical functionalities while providing information on biosynthesis, physiological context ...[more]

PMID: 32325029

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