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Examination of Galectin-3 Recruitment into Multivesicular Bodies for Exosomal Secretion

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Galectins

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2442))

Abstract

Cells use unconventional secretion to deliver the β-galactoside binding lectin galectin-3 from the cell interior into the extracellular milieu. This process starts with galectin-3 recruitment into intraluminal vesicles (ILVs), which are later released at the plasma membrane as exosomes. Electron microscopy is utilized to determine the location of GFP-tagged galectin-3 in pelleted exosomes. We also describe how these vesicles are harvested from cell culture media to determine their composition. The fluorescent protein GFP was fused with the exosomal sorting motif of galectin-3 to direct GFP into exosomes. Recruitment of this fusion construct into the lumen of exosomes can be assessed by proteinase K accessibility analysis.

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Authors and Affiliations

  1. Department of Cell Biology and Cell Pathology, Philipps University of Marburg, Marburg, Germany

    Sebastian Bänfer, Sophie Kutscher & Ralf Jacob

Authors
  1. Sebastian Bänfer
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  2. Sophie Kutscher
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  3. Ralf Jacob
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Corresponding authors

Correspondence to Sebastian Bänfer or Ralf Jacob .

Editor information

Editors and Affiliations

  1. Joint Program in Transfusion Medicine Brigham and Women’s Hospital, Harvard Medical School, Boston, MA, USA

    Sean R. Stowell

  2. Joint Program in Transfusion Medicine Brigham and Women’s Hospital, Harvard Medical School, Boston, MA, USA

    Connie M. Arthur

  3. Department of Surgery Beth Israel Deaconess Medical Center National Center for Functional Glycomics, Harvard Medical School, Boston, MA, USA

    Richard D. Cummings

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Bänfer, S., Kutscher, S., Jacob, R. (2022). Examination of Galectin-3 Recruitment into Multivesicular Bodies for Exosomal Secretion. In: Stowell, S.R., Arthur, C.M., Cummings, R.D. (eds) Galectins. Methods in Molecular Biology, vol 2442. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2055-7_22

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  • DOI: https://doi.org/10.1007/978-1-0716-2055-7_22

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2054-0

  • Online ISBN: 978-1-0716-2055-7

  • eBook Packages: Springer Protocols

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