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Molecular & Cellular Proteomics 4:191-204, 2005.
© 2005 by The American Society for Biochemistry and Molecular Biology, Inc.

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Research

Quantitative Proteomic Analysis of Sokotrasterol Sulfate-stimulated Primary Human Endothelial Cells ^*,S

Aly Karsan^,,¶,, Ingrid Pollet, Li-Rong Yu^¶, King C. Chan^¶, Thomas P. Conrads^¶, David A. Lucas^¶, Raymond Andersen^|| and Timothy Veenstra^¶

From the Department of Medical Biophysics and Pathology and Laboratory Medicine, British Columbia Cancer Agency, Vancouver, British Columbia V5Z 1L3, Canada, ^¶ Laboratory of Proteomics and Analytical Technologies, SAIC-Frederick, Inc., NCI, National Institutes of Health, Frederick, Maryland 21702-1201, and ^|| Department of Chemistry, University of British Columbia, Vancouver, British Columbia V6T 1Z4, Canada

The endothelium forms a continuous monolayer at the interface between blood and tissue and contributes significantly to the sensing and transducing of signals between blood and tissue. New blood vessel formation, or angiogenesis, is initiated by the activation of endothelial cells and is an important process required for various pathological and physiological situations. This study used cleavable isotope-coded affinity tag reagents combined with mass spectrometry to investigate the molecular basis of a recently discovered angiogenesis-promoting steroid, sokotrasterol sulfate. Changes in the relative abundances of over 1000 proteins within human endothelial cells treated with sokotrasterol sulfate and vehicle-treated cells were identified and quantitated using this technique. A method that examines the entire ensemble of quantitative measurements was developed to identify proteins that showed a statistically significant change in relative abundance resulting from treatment with sokotrasterol sulfate. A total of 93 proteins was significantly up-regulated, and 37 were down-regulated in response to sokotrasterol sulfate stimulation of endothelial cells. Among the up-regulated proteins, several were identified that are novel to endothelial cells and are likely involved in cell communication and morphogenesis. These findings are consistent with a role for sokotrasterol sulfate in endothelial sprouting.

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