Identification of the surface-accessible, lineage-specific vascular proteome by two-dimensional peptide mapping

FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology
Christoph RoesliMichael Detmar

Abstract

The formation of blood vessels (angiogenesis) and of lymphatic vessels (lymphangiogenesis) actively contributes to cancer progression and inflammation. Thus, there has been a quest for identifying the molecular mechanisms that control lymphatic and blood vessel formation and function. Membrane and extracellular matrix proteins can serve as suitable targets for imaging and/or therapeutic targeting; however, conventional proteomic technologies often fail to identify them systematically due to insolubility in water and low abundance of membrane proteins. To circumvent this problem, we applied a gel-free proteomics methodology termed two-dimensional peptide mapping (2D-PM) to cultured blood vascular (BECs) and lymphatic (LECs) endothelial cells. 2D-PM comprises biotinylation of surface-accessible proteins, their selective enrichment, separation by HPLC, and analysis by mass spectrometry. We identified 184 proteins that were specifically or predominantly expressed by LECs and 185 proteins specifically expressed by BECs, whereas 377 additional proteins were equally detected in both cell types. For representative proteins, the differential, lineage-specific expression was confirmed by Western analyses of cultured cells and by differen...Continue Reading

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Citations

Feb 24, 2012·Analytical and Bioanalytical Chemistry·Amelie S Benk, Christoph Roesli
Jul 24, 2008·Cell and Tissue Research·Giorgia Jurisic, Michael Detmar
Dec 20, 2008·Lymphatic Research and Biology·Leah N Cueni, Michael Detmar
Oct 27, 2012·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Natalie O KarpinichKathleen M Caron
Dec 4, 2012·International Journal of Molecular Sciences·Rocco SavinoRosa Terracciano
Mar 5, 2014·The Journal of Clinical Investigation·Sinem Karaman, Michael Detmar
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