CD109 is a component of exosome secreted from cultured cells

Hiroki Sakakura, Shinji Mii, Sumitaka Hagiwara, Takuya Kato, Noriyuki Yamamoto, Hideharu Hibi, Masahide Takahashi, Yoshiki Murakumo

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8 Citations (Scopus)

Abstract

Exosomes are 50-100-nm-diameter membrane vesicles released from various types of cells. Exosomes retain proteins, mRNAs and miRNAs, which can be transported to surrounding cells. CD109 is a glycosylphosphatidylinositol-anchored glycoprotein, and is released from the cell surface to the culture medium in vitro. Recently, it was reported that secreted CD109 from the cell surface downregulates transforming growth factor-β signaling in human keratinocytes. In this study, we revealed that CD109 is a component of the exosome in conditioned medium. FLAG-tagged human CD109 (FLAG-CD109) in conditioned medium secreted from HEK293 cells expressing FLAG-CD109 (293/FLAG-CD109) was immunoprecipitated with anti-FLAG affinity gel, and the co-precipitated proteins were analyzed by mass spectrometry and western blotting. Exosomal proteins were associated with CD109. We revealed the presence of CD109 in exosome fractions from conditioned medium of 293/FLAG-CD109. Moreover, the localization of CD109 in the exosome was demonstrated using immuno-electron microscopy. When we used HEK293 cells expressing FLAG-tagged truncated CD109, which does not contain the C-terminal region, the association of truncated CD109 with exosomes was not detected in conditioned medium. These findings indicate that CD109 is an exosomal protein and that the C-terminal region of CD109 is required for its presence in the exosome.

Original languageEnglish
Pages (from-to)816-822
Number of pages7
JournalBiochemical and Biophysical Research Communications
Volume469
Issue number4
DOIs
Publication statusPublished - 22-01-2016
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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  • Cite this

    Sakakura, H., Mii, S., Hagiwara, S., Kato, T., Yamamoto, N., Hibi, H., Takahashi, M., & Murakumo, Y. (2016). CD109 is a component of exosome secreted from cultured cells. Biochemical and Biophysical Research Communications, 469(4), 816-822. https://doi.org/10.1016/j.bbrc.2015.12.063