Proteomic analysis of vascular endothelial cells in response to laminar shear stress.
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 >or=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.
Wang
Xiao-Li
XL
Department of Internal Medicine, Mayo Clinic, Rochester, MN, USA.
Fu
Alex
A
Department of Radiology, Mayo Clinic, Rochester, MN, USA.
Raghavakaimal
Sreekumar
S
Department of Endocrinology, Mayo Clinic, Rochester, MN, USA.
Lee
Hon-Chi
HC
Department of Internal Medicine, Mayo Clinic, Rochester, MN, USA.
D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D002853:Chromatography, Liquid; D042783:Endothelial Cells / Q000502:physiology*; D018840:HSP70 Heat-Shock Proteins / Q000378:metabolism; D007553:Isotope Labeling; D020543:Proteome / Q000032:analysis*; D015398:Signal Transduction / Q000502:physiology; D013314:Stress, Mechanical; D053719:Tandem Mass Spectrometry
D018840:HSP70 Heat-Shock Proteins; D020543:Proteome