Expanding the repertoire of GalNAc analogues for cell-specific bioorthogonal tagging of glycoproteins.

Autor:	Zafar A; Chemical Glycobiology Laboratory, The Francis Crick Institute NW1 1AT London UK.; Department of Chemistry, Imperial College London W12 0BZ London UK b.schumann@imperial.ac.uk., Sridhar S; Chemical Glycobiology Laboratory, The Francis Crick Institute NW1 1AT London UK.; Department of Chemistry, Imperial College London W12 0BZ London UK b.schumann@imperial.ac.uk.; Tumour-Host Interaction Laboratory, The Francis Crick Institute NW1 1AT London UK., Bineva-Todd G; Chemical Glycobiology Laboratory, The Francis Crick Institute NW1 1AT London UK., Cioce A; Chemical Glycobiology Laboratory, The Francis Crick Institute NW1 1AT London UK.; Department of Chemistry, Imperial College London W12 0BZ London UK b.schumann@imperial.ac.uk., Abdulla N; Chemical Glycobiology Laboratory, The Francis Crick Institute NW1 1AT London UK.; Department of Chemistry, Imperial College London W12 0BZ London UK b.schumann@imperial.ac.uk., Chang V; Department of Chemistry, Yale University CT 06511 New Haven USA., Malaker SA; Department of Chemistry, Yale University CT 06511 New Haven USA., Hewings DS; Vertex Pharmaceuticals (Europe) Ltd., 86-88 Jubilee Avenue, Milton Park Abingdon OX14 4RW Oxfordshire UK., Schumann B; Chemical Glycobiology Laboratory, The Francis Crick Institute NW1 1AT London UK.; Department of Chemistry, Imperial College London W12 0BZ London UK b.schumann@imperial.ac.uk.
Jazyk:	angličtina
Zdroj:	RSC chemical biology [RSC Chem Biol] 2024 Aug 22. Date of Electronic Publication: 2024 Aug 22.
DOI:	10.1039/d4cb00093e
Abstrakt:	Glycosylation is a ubiquitous modification of proteins, necessitating approaches for its visualization and characterization. Bioorthogonally tagged monosaccharides have been instrumental to this end, offering a chemical view into the cell biology of glycans. Understanding the use of such monosaccharides by cellular biosynthetic pathways has expanded their applicability in cell biology, for instance through the strategy named Bio-Orthogonal Cell-specific TAgging of Glycoproteins (BOCTAG). Here, we show that the cellular use of two azide-tagged analogues of the monosaccharide N -acetylgalactosamine (GalNAzMe and GalNPrAz) can be promoted through expression of two biosynthetic enzymes. More precisely, cellular expression of the bacterial kinase NahK and the engineered human pyrophosphorylase AGX1 F383A led to biosynthesis of the corresponding activated nucleotide-sugars and subsequent bioorthogonal tagging of the cellular glycoproteome. We explore the use of both sugars for BOCTAG, demonstrating the visualization of cell surface glycosylation tagged with GalNPrAz in a specific cell line in a co-culture system. Our work adds to the toolbox of glycoprotein analysis in biomedicine. Competing Interests: S. A. M. is an inventor on a Stanford patent related to the use of mucinase digestion for glycoproteomic analysis and is a consultant for InterVenn Biosciences. (This journal is © The Royal Society of Chemistry.)
Databáze:	MEDLINE
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