Flow-induced Reorganization of Laminin-integrin Networks Within the Endothelial Basement Membrane Uncovered by Proteomics.

Autor:	Béguin EP; Department of Molecular and Cellular Hemostasis, Sanquin Research, Amsterdam, The Netherlands., Janssen EFJ; Department of Molecular and Cellular Hemostasis, Sanquin Research, Amsterdam, The Netherlands., Hoogenboezem M; Department of Molecular and Cellular Hemostasis, Sanquin Research, Amsterdam, The Netherlands., Meijer AB; Department of Molecular and Cellular Hemostasis, Sanquin Research, Amsterdam, The Netherlands; Department of Biomolecular Mass Spectrometry, Utrecht Institute for Pharmaceutical Sciences (UIPS), Utrecht University, Utrecht, The Netherlands., Hoogendijk AJ; Department of Molecular and Cellular Hemostasis, Sanquin Research, Amsterdam, The Netherlands., van den Biggelaar M; Department of Molecular and Cellular Hemostasis, Sanquin Research, Amsterdam, The Netherlands. Electronic address: m.vandenbiggelaar@sanquin.nl.
Jazyk:	angličtina
Zdroj:	Molecular & cellular proteomics : MCP [Mol Cell Proteomics] 2020 Jul; Vol. 19 (7), pp. 1179-1192. Date of Electronic Publication: 2020 Apr 24.
DOI:	10.1074/mcp.RA120.001964
Abstrakt:	The vessel wall is continuously exposed to hemodynamic forces generated by blood flow. Endothelial mechanosensors perceive and translate mechanical signals via cellular signaling pathways into biological processes that control endothelial development, phenotype and function. To assess the hemodynamic effects on the endothelium on a system-wide level, we applied a quantitative mass spectrometry approach combined with cell surface chemical footprinting. SILAC-labeled endothelial cells were subjected to flow-induced shear stress for 0, 24 or 48 h, followed by chemical labeling of surface proteins using a non-membrane permeable biotin label, and analysis of the whole proteome and the cell surface proteome by LC-MS/MS analysis. These studies revealed that of the >5000 quantified proteins 104 were altered, which were highly enriched for extracellular matrix proteins and proteins involved in cell-matrix adhesion. Cell surface proteomics indicated that LAMA4 was proteolytically processed upon flow-exposure, which corresponded to the decreased LAMA4 mass observed on immunoblot. Immunofluorescence microscopy studies highlighted that the endothelial basement membrane was drastically remodeled upon flow exposure. We observed a network-like pattern of LAMA4 and LAMA5, which corresponded to the localization of laminin-adhesion molecules ITGA6 and ITGB4. Furthermore, the adaptation to flow-exposure did not affect the inflammatory response to tumor necrosis factor α, indicating that inflammation and flow trigger fundamentally distinct endothelial signaling pathways with limited reciprocity and synergy. Taken together, this study uncovers the blood flow-induced remodeling of the basement membrane and stresses the importance of the subendothelial basement membrane in vascular homeostasis. Competing Interests: Conflict of interest—Authors declare no competing interests. (© 2020 Béguin et al.)
Databáze:	MEDLINE
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