Cell-specific bioorthogonal tagging of glycoproteins

Anna Cioce, Beatriz Calle, Tatiana Rizou, Sarah C. Lowery, Victoria L. Bridgeman, Keira E. Mahoney, Andrea Marchesi, Ganka Bineva-Todd, Helen Flynn, Zhen Li, Omur Y. Tastan, Chloe Roustan, Pablo Soro-Barrio, Mahmoud-Reza Rafiee, Acely Garza-Garcia, Aristotelis Antonopoulos, Thomas M. Wood, Tessa Keenan, Peter Both, Kun Huang, Fabio Parmeggian, Ambrosius P. Snijders, Mark Skehel, Svend Kjær, Martin A. Fascione, Carolyn R. Bertozzi, Stuart M. Haslam, Sabine L. Flitsch, Stacy A. Malaker, Ilaria Malanchi and Benjamin Schumann ()
Additional contact information
Anna Cioce: Imperial College London
Beatriz Calle: Imperial College London
Tatiana Rizou: The Francis Crick Institute
Sarah C. Lowery: Yale University
Victoria L. Bridgeman: The Francis Crick Institute
Keira E. Mahoney: Yale University
Andrea Marchesi: Imperial College London
Ganka Bineva-Todd: The Francis Crick Institute
Helen Flynn: The Francis Crick Institute
Zhen Li: Imperial College London
Omur Y. Tastan: The Francis Crick Institute
Chloe Roustan: The Francis Crick Institute
Pablo Soro-Barrio: The Francis Crick Institute
Mahmoud-Reza Rafiee: The Francis Crick Institute
Acely Garza-Garcia: The Francis Crick Institute
Aristotelis Antonopoulos: Imperial College London
Thomas M. Wood: Stanford University
Tessa Keenan: University of York
Peter Both: The University of Manchester
Kun Huang: The University of Manchester
Fabio Parmeggian: The University of Manchester
Ambrosius P. Snijders: The Francis Crick Institute
Mark Skehel: The Francis Crick Institute
Svend Kjær: The Francis Crick Institute
Martin A. Fascione: University of York
Carolyn R. Bertozzi: Stanford University
Stuart M. Haslam: Imperial College London
Sabine L. Flitsch: The University of Manchester
Stacy A. Malaker: Yale University
Ilaria Malanchi: The Francis Crick Institute
Benjamin Schumann: Imperial College London

Nature Communications, 2022, vol. 13, issue 1, 1-18

Abstract: Abstract Altered glycoprotein expression is an undisputed corollary of cancer development. Understanding these alterations is paramount but hampered by limitations underlying cellular model systems. For instance, the intricate interactions between tumour and host cannot be adequately recapitulated in monoculture of tumour-derived cell lines. More complex co-culture models usually rely on sorting procedures for proteome analyses and rarely capture the details of protein glycosylation. Here, we report a strategy termed Bio-Orthogonal Cell line-specific Tagging of Glycoproteins (BOCTAG). Cells are equipped by transfection with an artificial biosynthetic pathway that transforms bioorthogonally tagged sugars into the corresponding nucleotide-sugars. Only transfected cells incorporate bioorthogonal tags into glycoproteins in the presence of non-transfected cells. We employ BOCTAG as an imaging technique and to annotate cell-specific glycosylation sites in mass spectrometry-glycoproteomics. We demonstrate application in co-culture and mouse models, allowing for profiling of the glycoproteome as an important modulator of cellular function.

Date: 2022
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DOI: 10.1038/s41467-022-33854-0

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