Characterization of human vascular endothelial cadherin glycans

The glycosylation pattern of human vascular endothelial cad-herin (VE-cadherin), purified from cultured human umbili-cal cord vein endothelial cells, was analyzed. VE-cadherin was metabolically radiolabeled with D-[6-3H]glucosamine, isolated by immunoprecipitation, purified by SDS-PAGE and in-gel digested with endoproteinase Asp N. Oligosaccharides were sequentially released from resulting glycopeptides and analyzed by chromatographic profiling. The results revealed that VE-cadherin carries predominantly sialylated dianten-nary and hybrid-type glycans in addition to some trianten-nary and high mannose-type species. Highly branched, tetra-antennary oligosaccharides were found in trace amounts only. Immunohistochemical labeling of VE-cadherin and sialic acids displayed a codistribution along the intercellular junctions in endothelial cells of human umbilical arteries, veins, and cultured endothelial monolayers. Ca2+-depletion, performed on cultured endothelial cells, resulted in a revers-ible complete disappearance of VE-cadherin and of almost all sialic acid staining from the junctions. Sialidase treatment of whole cells caused a change of VE-cadherin immunofluores-cence from a continuous and netlike superstructural organiz-ation to a scattered inconsistent one. Hence, cell surface sialic acids might play a role in VE-cadherin organization. Key words: VE-cadherin/carbohydrate analysis/ oligosaccharides/VE-cadherin-superstructur