Comparative Glycomic Analysis of Exosome Subpopulations Derived from Pancreatic Cancer Cell Lines

Atsushi Matsuda, Atsushi Kuno, Maki Yoshida, Takanori Wagatsuma, Takashi Sato, Makoto Miyagishi, Jing Zhao, Makoto Suematsu, Yasuaki Kabe, Hisashi Narimatsu

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Extracellular vesicles such as exosomes are generally covered with an array of glycans, which are controlled by the host-cell glyco-synthetic machinery, similar to secreted and membrane glycoproteins. Several exosome subpopulations classified by their tetraspanin expression have been investigated in the context of diseases. However, a comparative analysis of their glycomics has never been attempted. Herein, we report a method for the comparative glycomic analysis of exosome subpopulations among pancreatic cancer cell lines. Glycomic profiles were obtained for extracellular vesicles, secreted glycoproteins, and membrane glycoproteins from eight cell lines. Statistical analyses revealed high populations of PHA-L-binding proteins in the vesicles. The surfaces of extracellular vesicles were labeled with Cy3 and captured by magnetic beads with antibodies against tetraspanins (CD9, CD63, and CD81). The coprecipitated vesicles were lysed and subjected to a lectin microarray analysis. A hierarchical clustering analysis using 19 glycomic profiles confirmed that most subpopulations, except CD81-positive exosomes, could be distinguished according to the host-cell species. Principal component analysis and subsequent lectin-affinity capturing of intact exosomes highlighted that CD81-positive exosomes preferentially expressed not PHA-L- but LEL-binding proteins on their surfaces. These data suggested that exosomal glycomics depended on the host-cell type and subpopulation.

Original languageEnglish
Pages (from-to)2516-2524
Number of pages9
JournalJournal of Proteome Research
Volume19
Issue number6
DOIs
Publication statusPublished - 2020 Jun 5

Keywords

  • exosome
  • glycomics
  • glycoprotein
  • lectin microarray

ASJC Scopus subject areas

  • Biochemistry
  • Chemistry(all)

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  • Cite this

    Matsuda, A., Kuno, A., Yoshida, M., Wagatsuma, T., Sato, T., Miyagishi, M., Zhao, J., Suematsu, M., Kabe, Y., & Narimatsu, H. (2020). Comparative Glycomic Analysis of Exosome Subpopulations Derived from Pancreatic Cancer Cell Lines. Journal of Proteome Research, 19(6), 2516-2524. https://doi.org/10.1021/acs.jproteome.0c00200