Identification of cell surface markers to differentiate rat endothelial and fibroblast cells using lectin arrays and LC-ESI-MS/MS.
Vascular endothelial cells located at the inner surface of blood vessels are a key component in angiogenesis and are employed as a primary cell type in the study of angiogenesis. These endothelial cells are, however, easily contaminated with fibroblast cells, which are located in proximity to the endothelial cells, during their isolation from tissue. It is thus important to find markers to distinguish the two cell types. In the present work, lectin arrays were prepared using aldehyde-terminated self-assembled monolayers (SAMs) and utilized to explore cell surface carbohydrate expression patterns on endothelial and fibroblast cells. It was found that the lectins Griffonia simplicifolia II (GS II) and Ulex europaeus agglutinin I (UEA I) selectively bind to rat fibroblast cells and not to rat endothelial cells. GS II-binding glycoproteins on fibroblast cells, which are potential cell surface markers to differentiate endothelial and fibroblast cells, were captured on a GS II lectin column and analyzed by LC-ESI-MS/MS. Six candidate cell surface glycoproteins were identified. Differential expression was confirmed by Western blot analysis for two of these proteins, lysosome-associated membrane glycoprotein-1 and transmembrane glycoprotein NMB.
Lee
Ji Eun
JE
Department of Chemistry, University of Wisconsin-Madison, 1101 University Avenue, Madison, Wisconsin 53706, USA.
Mirza
Shama P
SP
Didier
Daniela N
DN
Scalf
Mark
M
Olivier
Michael
M
Greene
Andrew S
AS
Smith
Lloyd M
LM
D000818:Animals; D015415:Biomarkers / Q000032:analysis; D002241:Carbohydrates / Q000032:analysis; D002462:Cell Membrane / Q000378:metabolism*; D002478:Cells, Cultured; D002853:Chromatography, Liquid / Q000379:methods*; D042783:Endothelial Cells / Q000378:metabolism*; D005347:Fibroblasts; D037102:Lectins / Q000032:analysis*; D040081:Protein Array Analysis / Q000379:methods*; D051381:Rats; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods*
D015415:Biomarkers; D002241:Carbohydrates; D037102:Lectins