In-depth proteomics identifies a role for autophagy in controlling reactive oxygen species mediated endothelial permeability
Authors list
Francesca Patella Lisa J Neilson Dimitris Athineos Zahra Erami Kurt Anderson Karen Blyth Kevin M Ryan Sara ZanivanAbstract
Endothelial cells (ECs) form the inner layer of blood vessels and physically separate the blood from the surrounding tissue. To support tissues with nutrients and oxygen, the endothelial monolayer is semipermeable. When EC permeability is altered, blood vessels are not functional, and this is associated with disease. A comprehensive knowledge of the mechanisms regulating EC permeability is key in developing strategies to target this mechanism in pathologies. Here we have used an in vitro model of human umbilical vein endothelial cells mimicking the formation of a physiologically permeable vessel and performed time-resolved in-depth molecular profiling using stable isotope labeling by amino acids in cell culture mass spectrometry (MS)-proteomics. Autophagy is induced when ECs are assembled into a physiologically permeable monolayer. By using siRNA and drug treatment to block autophagy in combination with functional assays and MS proteomics, we show that ECs require autophagy flux to maintain intracellular reactive oxygen species levels, and this is required to maintain the physiological permeability of the cells.
Journal details
Journal Journal of Proteome Research
Volume 15
Issue number 7
Pages 2187-2197
Available online
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Publisher website (DOI) 10.1021/acs.jproteome.6b00166
Europe PubMed Central 27246970
Pubmed 27246970
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