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Lipidomic and proteomic analysis of exosomes from mouse cortical collecting duct cells
Author(s) -
Dang Viet D.,
Jella Kishore Kumar,
Ragheb Ragy R. T.,
Denslow Nancy D.,
Alli Abdel A.
Publication year - 2017
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fj.201700417r
Subject(s) - microvesicles , sphingomyelin , microvesicle , microbiology and biotechnology , exosome , chemistry , annexin , basigin , annexin a2 , cardiolipin , membrane protein , biology , biochemistry , phospholipid , membrane , microrna , cell , matrix metalloproteinase , gene
Exosomes are endosome‐derived nanovesicles that are involved in cellular communication and signaling. Exosomes are produced by epithelial cells and are found in biologic fluids including blood and urine. The packaged material within exosomes includes proteins and lipids, but the molecular comparison within exosome subtypes is largely unknown. The purpose of this study was to investigate differences between exosomes derived from the apical plasma membrane and basolateral plasma membrane of polarized murine cortical collecting duct principal cells. Nanoparticle tracking analysis showed that the size and concentration of apical and basolateral exosomes remained relatively stable across 3 different temperatures (23, 37, and 42°C). Liquid chromatography–tandem mass spectrometry analysis revealed marked differences between the proteins packaged within the two types of exosomes from the same cells. Several proteins expressed at the inner leaflet of the plasma membrane, including α‐actinin‐1, moesin, 14‐3‐3 protein ζ/δ, annexin A1/A3/A4/A5/A6, clathrin heavy chain 1, glyceraldehyde‐3‐phosphate dehydrogenase, α‐enolase, filamin‐A, and heat shock protein 90, were identified in samples of apical plasma membrane‐derived exosomes, but not in basolateral plasma membrane exosomes from mouse cortical collecting duct cells. In addition to differences at the protein level, mass spectrometry–based shotgun lipidomics analysis showed significant differences in the lipid classes and fatty acid composition of the two types of exosomes. We found higher levels of sphingomyelin and lower levels of cardiolipin, among other phospholipids in the apical plasma membrane compared to the basolateral plasma membrane exosomes. The molecular analyses of exosome subtypes presented herein will contribute to our understanding of exosome biogenesis, and the results may have potential implications for biomarker discovery.—Dang, V. D., Jella, K. K., Ragheb, R. R. T., Denslow, N. D., Alli, A. A. Lipidomic and proteomic analysis of exosomes from mouse cortical collecting duct cells. FASEB J. 31, 5399–5408 (2017). www.fasebj.org