Demonstration of single chain urokinase-type plasminogen activator on human platelet membrane.
Fibrinolytic activity was found to be associated with sonicated platelet membranes after separation from cytosol by differential centrifugation. This fibrinolytic activity was attributed to the presence of a plasminogen activator, which was immunochemically identified as urinary-type plasminogen activator (uPA) by antibody neutralization assay, immunoblotting, and immunofluorescence. The molecular weight (mol wt) of this uPA was 54,000 and was present as the single chain form, although a small amount was detected in a higher mol wt complex indicative of a uPA-inhibitor complex. Treatment of membrane preparations with Triton X-100, 3 mol/L KCl, and 0.1 mol/L glycine, (pH 2.3), but not 10 mmol/L ethylenediamine tetraacetic acid (EDTA), removed the uPA from the membrane. This suggests that uPA is a peripheral membrane protein and that metal ions do not mediate protein-membrane association. Immunofluorescent staining revealed the presence of uPA on the outer surface of the platelet in preparations of intact unstimulated platelets. Thus, uPA is associated with the outer leaflet of the platelet membrane and may be involved with the acceleration of thrombus degradation observed with platelet-rich thrombi.
Park
S
S
Department of Basic and Clinical Research, Scripps Clinic and Research Foundation, La Jolla, CA 92037.
Harker
L A
LA
Marzec
U M
UM
Levin
E G
EG
D001792:Blood Platelets / Q000201:enzymology*; D015153:Blotting, Western; D002462:Cell Membrane / Q000201:enzymology; D005455:Fluorescent Antibody Technique; D006801:Humans; D008970:Molecular Weight; D013347:Subcellular Fractions / Q000201:enzymology; D014568:Urokinase-Type Plasminogen Activator / Q000097:blood*
D014568:Urokinase-Type Plasminogen Activator