The Francis Crick Institute
s41467-022-33854-0.pdf (2.19 MB)

Cell-specific bioorthogonal tagging of glycoproteins.

Download (2.19 MB)
journal contribution
posted on 2022-10-31, 12:56 authored by 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, Benjamin Schumann
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.


Crick (Grant ID: CC1068, Grant title: STP Structural Biology) Crick (Grant ID: CC1063, Grant title: STP Proteomics) Crick (Grant ID: CC1107, Grant title: STP Bioinformatics & Biostatistics) Crick (Grant ID: 10749, Grant title: Schumann FC001749) Crick (Grant ID: 10112, Grant title: Malanchi FC001112) Crick (Grant ID: 10060, Grant title: Carvalho FC001060)