Proteomic analysis of vascular endothelial cells in response to laminar shear stress

• Published in: Proteomics (Weinheim) Vol. 7; no. 4; pp. 588 - 596
• Main Authors: Wang, Xiao-Li, Fu, Alex, Raghavakaimal, Sreekumar, Lee, Hon-Chi
• Format: Journal Article
• Language: English
• Published: Weinheim WILEY-VCH Verlag 01.02.2007; WILEY‐VCH Verlag; Wiley-VCH
• Subjects: Analytical, structural and metabolic biochemistry; Animals; Biological and medical sciences; Cattle; Cells, Cultured; Chromatography, Liquid; Endothelial Cells - physiology; Endothelium; Fundamental and applied biological sciences. Psychology; HSP70 Heat-Shock Proteins - metabolism; Isotope Labeling; Isotope-coded affinity tag; Miscellaneous; Proteins; Proteome - analysis; Shear stress; Signal Transduction - physiology; Stress, Mechanical; Tandem Mass Spectrometry; Tandem MS; tope-coded affinity tag / Shear stress / Tandem MS; Endothelial cell; Stress; Proteomics; Endothelium
• ISSN: 1615-9853; 1615-9861
• DOI: 10.1002/pmic.200600568

Isotope‐coded affinity tags (cICAT) coupled with mass spectrometric analysis is one of the leading technologies for quantitative proteomic profiling and protein quantification. We performed proteomic analysis of bovine aortic endothelial cells (BAEC) in response to laminar shear stress using cICAT labeling coupled with LC‐MS/MS. Protein expressions in BAEC under 15 dynes/cm2 of shear stress for 10 min, 3 h, and 6 h were compared with matched stationary controls. Analysis of each sample produced 1800–2400 proteins at ≥75% confidence level. We found 142, 213, and 186 candidate proteins that were up‐ or down‐regulated by at least two‐fold after 10 min, 3 h, and 6 h of shear stress, respectively. Some of these proteins have known cellular functions and they encompass many signaling pathways. The signaling pathways that respond to shear stress include those of integrins, G‐protein‐coupled receptors, glutamate receptors, PI3K/AKT, apoptosis, Notch and cAMP‐mediated signaling pathways. The validity of the mass spectrometric analysis was also confirmed by Western blot and confocal immunofluorescence microscopy. The present quantitative proteomic analysis suggests novel potential regulatory mechanisms in vascular endothelial cells in response to shear stress. These results provide preliminary footprints for further studies on the signaling mechanisms induced by shear stress.