Aminooxy Biotin-Based Characterization of the Surfaceome of Chondrogenic Cells
Kovács, Patrik; Boocock, David J; Coveney, Clare; Mobasheri, Ali; Matta, Csaba (2025-05-01)
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Sisältö avataan julkiseksi: 01.05.2026
Humana press
01.05.2025
Kovács, P., Boocock, D.J., Coveney, C., Mobasheri, A., Matta, C. (2025). Aminooxy Biotin-Based Characterization of the Surfaceome of Chondrogenic Cells. In: Matta, C., Mobasheri, A., Takács, R. (eds) The Surfaceome. Methods in Molecular Biology, vol 2908. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-4434-8_5
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© 2025 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature
https://rightsstatements.org/vocab/InC/1.0/
© 2025 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature
https://rightsstatements.org/vocab/InC/1.0/
Julkaisun pysyvä osoite on
https://urn.fi/URN:NBN:fi:oulu-202505123287
https://urn.fi/URN:NBN:fi:oulu-202505123287
Tiivistelmä
Abstract
This protocol outlines a comprehensive approach for characterizing the cell surface subproteome of chondroprogenitor cells, based on aminooxy biotinylation followed by mass spectrometry analysis. The first step involves the selective labeling of cell surface proteins with aminooxy biotin on living chondroprogenitor cells, ensuring the specific tagging of glycoproteins on the outer membrane. Subsequently, glycocapture technique is employed to enrich the glycosylated fraction of the cell surface proteins. Following multiple wash steps to reduce contamination with detergents and nonsurface proteins, shotgun mass spectrometry is applied for the quantitative and qualitative analysis of the enriched subproteome, allowing for the identification and characterization of surface proteins. The integration of these techniques offers a comprehensive and sensitive method for profiling the cell surface proteome during chondrogenesis, enabling a deeper understanding of the molecular composition of chondroprogenitor cells. This protocol holds promise for advancing our knowledge of chondrogenesis and may contribute to the identification of potential therapeutic targets for cartilage-related disorders.
This protocol outlines a comprehensive approach for characterizing the cell surface subproteome of chondroprogenitor cells, based on aminooxy biotinylation followed by mass spectrometry analysis. The first step involves the selective labeling of cell surface proteins with aminooxy biotin on living chondroprogenitor cells, ensuring the specific tagging of glycoproteins on the outer membrane. Subsequently, glycocapture technique is employed to enrich the glycosylated fraction of the cell surface proteins. Following multiple wash steps to reduce contamination with detergents and nonsurface proteins, shotgun mass spectrometry is applied for the quantitative and qualitative analysis of the enriched subproteome, allowing for the identification and characterization of surface proteins. The integration of these techniques offers a comprehensive and sensitive method for profiling the cell surface proteome during chondrogenesis, enabling a deeper understanding of the molecular composition of chondroprogenitor cells. This protocol holds promise for advancing our knowledge of chondrogenesis and may contribute to the identification of potential therapeutic targets for cartilage-related disorders.
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