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Detailed Analysis o...
Detailed Analysis of Protein Topology of Extracellular Vesicles–Evidence of Unconventional Membrane Protein Orientation
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- Cvjetkovic, Aleksander (author)
- Gothenburg University,Göteborgs universitet,Krefting Research Centre
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- Jang, Su Chul, 1984 (author)
- Gothenburg University,Göteborgs universitet,Krefting Research Centre
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- Konečná, Barbora (author)
- Gothenburg University,Göteborgs universitet,Krefting Research Centre
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- Höög, Johanna L, 1979 (author)
- Gothenburg University,Göteborgs universitet,Krefting Research Centre,Institutionen för kemi och molekylärbiologi,Department of Chemistry and Molecular Biology
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- Sihlbom, Carina, 1973 (author)
- Gothenburg University,Göteborgs universitet,Core Facilities, Proteomics,Core Facilities, Proteomics
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- Lässer, Cecilia, 1981 (author)
- Gothenburg University,Göteborgs universitet,Krefting Research Centre
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- Lötvall, Jan, 1956 (author)
- Gothenburg University,Göteborgs universitet,Krefting Research Centre
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(creator_code:org_t)
- 2016-11-08
- 2016
- English.
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In: Scientific Reports. - : Springer Science and Business Media LLC. - 2045-2322. ; 6
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Abstract
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- Extracellular vesicles (EVs) are important mediators of intercellular communication that change the recipient cell by shuttling lipids, RNA, or protein cargo between cells. Here, we investigate the topology of the protein cargo found in EVs, as this topology can fundamentally influence the biological effects of EVs. A multiple proteomics approach, combining proteinase treatment and biotin tagging, shows that many proteins of cytosolic origin are localized on the surface of EVs. A detailed analysis of the EV proteome at the peptide level revealed that a number of EV membrane proteins are present in a topologically reversed orientation compared to what is annotated. Two examples of such proteins, SCAMP3 and STX4, were confirmed to have a reversed topology. This reversed typology was determined using flow cytometry and fluorescent microscopy with antibodies directed toward their cytoplasmic epitopes. These results describe a novel workflow to define the EV proteome and the orientation of each protein, including membrane protein topology. These data are fundamentally important to understanding the EV proteome and required to fully explain EV biogenesis as well as biological function in recipient cells.
Subject headings
- NATURVETENSKAP -- Biologi -- Cellbiologi (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences -- Cell Biology (hsv//eng)
Keyword
- exosome
- proteomics
- electron microscopy
- vesicle
Publication and Content Type
- ref (subject category)
- art (subject category)
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- By the author/editor
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Cvjetkovic, Alek ...
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Jang, Su Chul, 1 ...
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Konečná, Barbora
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Höög, Johanna L, ...
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Sihlbom, Carina, ...
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Lässer, Cecilia, ...
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Lötvall, Jan, 19 ...
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- About the subject
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- NATURAL SCIENCES
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NATURAL SCIENCES
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and Biological Scien ...
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and Cell Biology
- Articles in the publication
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Scientific Repor ...
- By the university
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University of Gothenburg