Affiliations 

  • 1 Department of Neurology, Massachusetts General Hospital, Charlestown, MA, 02129, USA. mikolaj.zaborowski@gmail.com
  • 2 Department of Neurology, Massachusetts General Hospital, Charlestown, MA, 02129, USA
  • 3 Center for Systems Biology, Massachusetts General Hospital, Boston, MA, 02114, USA
  • 4 Department of Medicine, Vanderbilt University Medical Center, Nashville, TN, 37232, USA
  • 5 Department of Neurology, Massachusetts General Hospital, Charlestown, MA, 02129, USA. breakefield@hms.harvard.edu
Sci Rep, 2019 11 22;9(1):17387.
PMID: 31758005 DOI: 10.1038/s41598-019-53554-y

Abstract

Extracellular vesicles (EVs) released by cells play a role in intercellular communication. Reporter and targeting proteins can be modified and exposed on the surface of EVs to investigate their half-life and biodistribution. A characterization of membrane-bound Gaussia luciferase (mbGluc) revealed that its signal was detected also in a form smaller than common EVs (<70 nm). We demonstrated that mbGluc initially exposed on the surface of EVs, likely undergoes proteolytic cleavage and processed fragments of the protein are released into the extracellular space in active form. Based on this observation, we developed a new assay to quantitatively track shedding of membrane proteins from the surface of EVs. We used this assay to show that ectodomain shedding in EVs is continuous and is mediated by specific proteases, e.g. metalloproteinases. Here, we present a novel tool to study membrane protein cleavage and release using both in vitro and in vivo models.

* Title and MeSH Headings from MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.