Urokinase receptors in human monocytes.
Receptors for the 54 kDa plasminogen activator urokinase were characterized in freshly isolated and 5-14 day cultured human monocytes. The half saturation constant was about 55 pM in freshly isolated monocytes at 4 degrees C and 140 pM at 37 degrees C. Diisopropylfluorophosphate-inactivated urokinase was bound with the same affinity as catalytically active urokinase. Binding per cell of 2-5 pM urokinase increased progressively during cell culture with a concomitant decrease in the apparent affinity. By 14 days, binding had increased 5-7-fold and the half-saturation constant had increased to 500 pM at 4 degrees C, indicating a large increase in the binding capacity. Affinity cross-linking of labelled urokinase to receptors showed a 110 kDa complex in both freshly isolated and cultured monocytes. When cells with labelled urokinase (prebound at 4 degrees C) were incubated at 37 degrees C, about 80% of the urokinase dissociated as the intact molecule, whereas about 20% was degraded to iodide and iodotyrosine. Electron microscopic autoradiography of cultured monocytes incubated at 4 degrees C showed a marked heterogeneity between cells with regard to bound urokinase. Autoradiographic grains were mainly seen over the plasma membrane in areas rich in microvilli and invaginations. Transfer of the cells to 37 degrees C caused no major alteration in the distribution of grains. Thus, freshly prepared monocytes have urokinase receptors (approx. 55 kDa) of high affinity. Development to macrophage-like cells in culture causes a decrease in affinity and a large increase in capacity. The receptors are confined mainly to certain areas of the plasma membrane. Internalization and degradation of the ligand occurs only to a minor extent.
Nykjaer
A
A
Institute of Physiology, University of Aarhus, Denmark.
Petersen
C M
CM
Christensen
E I
EI
Davidsen
O
O
Gliemann
J
J
D003432:Cross-Linking Reagents / Q000378:metabolism; D006801:Humans; D007531:Isoflurophate / Q000378:metabolism; D007700:Kinetics; D009000:Monocytes / Q000201:enzymology / Q000378:metabolism* / Q000648:ultrastructure; D010960:Plasminogen Activators; D011956:Receptors, Cell Surface / Q000378:metabolism* / Q000648:ultrastructure; D055293:Receptors, Urokinase Plasminogen Activator; D014407:Tumor Cells, Cultured; D014568:Urokinase-Type Plasminogen Activator / Q000378:metabolism*
D003432:Cross-Linking Reagents; C524898:PLAUR protein, human; D011956:Receptors, Cell Surface; D055293:Receptors, Urokinase Plasminogen Activator; D007531:Isoflurophate; D010960:Plasminogen Activators; D014568:Urokinase-Type Plasminogen Activator