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Cell-specific bioorthogonal tagging of glycoproteins.

Nature communications | 2022

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.

Pubmed ID: 36284108 RIS Download

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Associated grants

  • Agency: Biotechnology and Biological Sciences Research Council, United Kingdom
    Id: EP/S005226/1
  • Agency: NCI NIH HHS, United States
    Id: R01 CA200423
  • Agency: NIGMS NIH HHS, United States
    Id: R35 GM147039
  • Agency: Biotechnology and Biological Sciences Research Council, United Kingdom
    Id: BB/T01279X/1
  • Agency: Biotechnology and Biological Sciences Research Council, United Kingdom
    Id: BB/M028836/1
  • Agency: Biotechnology and Biological Sciences Research Council, United Kingdom
    Id: BB/M02847X/1
  • Agency: Biotechnology and Biological Sciences Research Council, United Kingdom
    Id: BB/M027791/1
  • Agency: Biotechnology and Biological Sciences Research Council, United Kingdom
    Id: EP/S013741/1
  • Agency: Wellcome Trust, United Kingdom
    Id: 218304/Z/19/Z

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K-562 (tool)

RRID:CVCL_0004

Cell line K-562 is a Cancer cell line with a species of origin Homo sapiens (Human)

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MCF-7 (tool)

RRID:CVCL_0031

Cell line MCF-7 is a Cancer cell line with a species of origin Homo sapiens (Human)

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