Cell-specific bioorthogonal tagging of glycoproteins

Anna  Anna; Beatriz Calle; Tatiana Rizou; Sarah Lowery; Victoria Bridgeman; Keira Mahoney; Andrea Marchesi; Ganka Bineva-Todd; Helen Flynn; Zhen Li; Omur Yilmaz Tastan; Chloe Roustan; Pablo  Soro-Barrio; Mahmoud-Reza Rafiee; Acely Garza-Garcia; Aristotelis Antonopoulos; Thomas Wood; Tessa Keenan; Peter Both; Kun Huang; Fabio Parmeggiani; Ambrosius Snijders; John Skehel; Svend Kjaer; Martin Anthony Fascione; Carolyn R. Bertozzi; Stacey Malaker; Ilaria Malanchi; Benjamin Schumann

doi:10.1038/s41467-022-33854-0

Cell-specific bioorthogonal tagging of glycoproteins

Anna Anna , Beatriz Calle, Tatiana Rizou, Sarah Lowery, Victoria Bridgeman, Keira Mahoney, Andrea Marchesi, Ganka Bineva-Todd, Helen Flynn, Zhen Li, Omur Yilmaz Tastan, Chloe Roustan, Pablo Soro-Barrio, Mahmoud-Reza Rafiee, Acely Garza-Garcia, Aristotelis Antonopoulos, Thomas Wood, Tessa Keenan, Peter Both, Kun HuangFabio Parmeggiani, Ambrosius Snijders, John Skehel, Svend Kjaer, Martin Anthony Fascione, Carolyn R. Bertozzi, Stacey Malaker, Ilaria Malanchi, Benjamin Schumann^*

^*Corresponding author for this work

Chemistry

Research output: Contribution to journal › Article › peer-review

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.

Original language	English
Article number	6237
Number of pages	18
Journal	Nature Communications
Volume	13
DOIs	https://doi.org/10.1038/s41467-022-33854-0
Publication status	Published - 25 Oct 2022

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10.1038/s41467-022-33854-0Licence: CC BY

s41467-022-33854-0
© The Author(s) 2022
Final published version, 2.19 MBLicence: CC BY

Chemo-enzymatic Production of Specialty Glycans
Fascione, M. A.
19/10/15 → 18/10/20
Project: Research project (funded) › Research

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Anna , A., Calle, B., Rizou, T., Lowery, S., Bridgeman, V., Mahoney, K., Marchesi, A., Bineva-Todd, G., Flynn, H., Li, Z., Yilmaz Tastan, O., Roustan, C., Soro-Barrio, P., Rafiee, M.-R., Garza-Garcia, A., Antonopoulos, A., Wood, T., Keenan, T., Both, P., ... Schumann, B. (2022). Cell-specific bioorthogonal tagging of glycoproteins. Nature Communications, 13, Article 6237. https://doi.org/10.1038/s41467-022-33854-0

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title = "Cell-specific bioorthogonal tagging of glycoproteins",

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.",

author = "Anna Anna and Beatriz Calle and Tatiana Rizou and Sarah Lowery and Victoria Bridgeman and Keira Mahoney and Andrea Marchesi and Ganka Bineva-Todd and Helen Flynn and Zhen Li and {Yilmaz Tastan}, Omur and Chloe Roustan and Pablo Soro-Barrio and Mahmoud-Reza Rafiee and Acely Garza-Garcia and Aristotelis Antonopoulos and Thomas Wood and Tessa Keenan and Peter Both and Kun Huang and Fabio Parmeggiani and Ambrosius Snijders and John Skehel and Svend Kjaer and Fascione, {Martin Anthony} and Bertozzi, {Carolyn R.} and Stacey Malaker and Ilaria Malanchi and Benjamin Schumann",

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doi = "10.1038/s41467-022-33854-0",

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Anna , A, Calle, B, Rizou, T, Lowery, S, Bridgeman, V, Mahoney, K, Marchesi, A, Bineva-Todd, G, Flynn, H, Li, Z, Yilmaz Tastan, O, Roustan, C, Soro-Barrio, P, Rafiee, M-R, Garza-Garcia, A, Antonopoulos, A, Wood, T, Keenan, T, Both, P, Huang, K, Parmeggiani, F, Snijders, A, Skehel, J, Kjaer, S, Fascione, MA, Bertozzi, CR, Malaker, S, Malanchi, I & Schumann, B 2022, 'Cell-specific bioorthogonal tagging of glycoproteins', Nature Communications, vol. 13, 6237. https://doi.org/10.1038/s41467-022-33854-0

TY - JOUR

T1 - Cell-specific bioorthogonal tagging of glycoproteins

AU - Anna , Anna

AU - Calle, Beatriz

AU - Rizou, Tatiana

AU - Lowery, Sarah

AU - Bridgeman, Victoria

AU - Mahoney, Keira

AU - Marchesi, Andrea

AU - Bineva-Todd, Ganka

AU - Flynn, Helen

AU - Li, Zhen

AU - Yilmaz Tastan, Omur

AU - Roustan, Chloe

AU - Soro-Barrio, Pablo

AU - Rafiee, Mahmoud-Reza

AU - Garza-Garcia, Acely

AU - Antonopoulos, Aristotelis

AU - Wood, Thomas

AU - Keenan, Tessa

AU - Both, Peter

AU - Huang, Kun

AU - Parmeggiani, Fabio

AU - Snijders, Ambrosius

AU - Skehel, John

AU - Kjaer, Svend

AU - Fascione, Martin Anthony

AU - Bertozzi, Carolyn R.

AU - Malaker, Stacey

AU - Malanchi, Ilaria

AU - Schumann, Benjamin

PY - 2022/10/25

Y1 - 2022/10/25

N2 - 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.

AB - 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.

U2 - 10.1038/s41467-022-33854-0

DO - 10.1038/s41467-022-33854-0

M3 - Article

SN - 2041-1723

VL - 13

JO - Nature Communications

JF - Nature Communications

M1 - 6237

ER -

Cell-specific bioorthogonal tagging of glycoproteins

Abstract

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Projects

Chemo-enzymatic Production of Specialty Glycans

Cite this