Identification of an endothelial cell surface protein that binds plasminogen.
To identify and characterize endothelial cell surface components that bind plasminogen, we used ligand-blotting to study binding of plasminogen to sodium dodecyl sulphate solubilized extracts of human umbilical vein endothelial cells. It was observed that glu-plasminogen bound predominantly to a 45 kDa endothelial cell polypeptide. The interaction of labelled glu-plasminogen with this polypeptide was reversible and specific as the binding could be inhibited by both excess cold lysine and unlabelled glu-plasminogen but not by unrelated proteins. Binding of glu-plasminogen to cell extracts prepared from endothelial cells that had been pretreated with proteinase K was significantly reduced indicating that the 45 kDa polypeptide is a cell-surface protein. The cell-surface localization of the 45 kDa polypeptide was also indicated by the positive interaction of glu-plasminogen with membrane fractions of endothelial cells. Lys-plasminogen also interacted with the 45 kDa polypeptide in a specific manner and reversibility experiments indicated that lys-plasminogen could also displace the bound glu-plasminogen. Since binding of plasminogen to the 45 kDa endothelial cell surface polypeptide was very similar to plasminogen binding to intact endothelial cells, we propose that the 45 kDa protein represents one of the major receptors for plasminogen on human endothelial cells.
Dudani
A K
AK
Ottawa Blood Centre, Canadian Red Cross Society, Ontario.
Hashemi
S
S
Aye
M T
MT
Ganz
P R
PR
D002478:Cells, Cultured; D004586:Electrophoresis; D019286:Endopeptidase K; D004730:Endothelium, Vascular / Q000737:chemistry* / Q000166:cytology / Q000378:metabolism; D006801:Humans; D008565:Membrane Proteins / Q000032:analysis* / Q000378:metabolism; D008970:Molecular Weight; D010958:Plasminogen / Q000378:metabolism*; D011956:Receptors, Cell Surface / Q000032:analysis* / Q000378:metabolism; D055293:Receptors, Urokinase Plasminogen Activator; D012697:Serine Endopeptidases / Q000378:metabolism
D008565:Membrane Proteins; C524898:PLAUR protein, human; D011956:Receptors, Cell Surface; D055293:Receptors, Urokinase Plasminogen Activator; D010958:Plasminogen; D012697:Serine Endopeptidases; D019286:Endopeptidase K