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Analysis of shear stress responsive proteins and signaling pathways in vascular endothelial cells by proteomic approach
Author(s) -
Wang XiaoLi,
Fu Alex,
Raghavakaimal Sreekumar,
Lee HonChi
Publication year - 2007
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/fasebj.21.6.a1432-d
Subject(s) - microbiology and biotechnology , shear stress , western blot , signal transduction , integrin , chemistry , receptor , proteome , endothelial stem cell , biology , biophysics , biochemistry , materials science , in vitro , composite material , gene
Laminar shear stress is one of the most important stimuli for vascular endothelial cell function. We performed proteomic analysis of bovine aortic endothelial cells (BAEC) in response to laminar shear stress using cICAT‐labeling coupled with liquid chromatography tandem mass spectrometry (LC‐MS/MS). Expression of proteins in BAEC subjected to shear stress of 15 dynes/cm 2 in a flow loop perfusion apparatus for 10 min, 3 h and 6 h were compared with matched stationary controls. Analysis of each sample identified 1,800–2,400 proteins at ≥75% confidence level. 142, 213, and 186 candidate proteins were found to have at least a 2‐fold up‐ or down‐regulation by 10 min, 3 h and 6 h of shear stress respectively. Some of these proteins have known cellular functions and they encompass many pathways including those of nitric oxide, integrins, insulin receptor, G‐protein‐coupled receptors, apoptosis, Notch and cAMP‐mediated signaling. The temporal pattern of changes varied among different signaling pathways. The validity of the mass spectrometric analysis was confirmed by Western blot and confocal immunofluorescence microscopy. This study suggests potential novel mechanisms in shear stress‐mediated vascular endothelial cell regulation. Hence, quantitative proteomic analysis is a promising approach for elucidating cellular protein regulation in response to physiological stimuli such as shear stress.