Plasminogen binding to rat hepatocytes in primary culture and to thin slices of rat liver.
Human 125I-plasminogen bound readily to rat hepatocytes in primary culture at 4 degrees C and at 37 degrees C. Binding was inhibited by lysine and reversed by lysine, epsilon-aminocaproic acid, or nonradiolabeled plasminogen. The Kd for binding of 125I-plasminogen to hepatocytes was 0.59 +/- 0.16 mumol/L, as determined from the saturation isotherm by nonlinear regression (r2 = 0.99) and the Scatchard transformation by linear regression (r2 = 0.93). The number of sites per cell was 14.1 +/- 1.1 x 10(6). Fibrinogen synthesis and secretion by hepatocytes was insufficient to account for the major fraction of plasminogen binding, as determined by enzyme-linked immunosorbent assay (ELISA). Polyacrylamide gel electrophoresis and trichloroacetic acid precipitation studies demonstrated that plasminogen is neither activated nor degraded when bound to hepatocytes at 37 degrees C. Thin slices of whole rat liver (500 microns), isolated and prepared totally at 4 degrees C, bound 125I-plasminogen. Binding was inhibited by lysine. 125I-albumin binding to liver slices was minimal and not inhibited by lysine. Activation of plasminogen by tissue plasminogen activator (t-PA) was enhanced by hepatocytes in primary culture. When lysine was included in the media, the enhanced rate of activation was no longer observed. After activation with t-PA, much of the plasmin remained associated with hepatocyte surfaces and was partially protected from inhibition by alpha 2-antiplasmin. These studies suggest that hepatocyte plasminogen binding sites may provide important surface anticoagulant activity.
Gonias
S L
SL
Department of Pathology, University of Virginia Health Sciences Center, Charlottesville 22908.
Braud
L L
LL
Geary
W A
WA
VandenBerg
S R
SR
D000818:Animals; D002478:Cells, Cultured; D005260:Female; D005340:Fibrinogen / Q000096:biosynthesis / Q000502:physiology; D005341:Fibrinolysin / Q000037:antagonists & inhibitors; D006801:Humans; D066298:In Vitro Techniques; D008099:Liver / Q000378:metabolism*; D010958:Plasminogen / Q000378:metabolism*; D011485:Protein Binding; D051381:Rats; D011919:Rats, Inbred Strains; D011956:Receptors, Cell Surface / Q000032:analysis*; D055293:Receptors, Urokinase Plasminogen Activator; D012711:Serum Albumin, Radio-Iodinated / Q000378:metabolism; D010959:Tissue Plasminogen Activator
C524898:PLAUR protein, human; C524900:Plaur protein, rat; D011956:Receptors, Cell Surface; D055293:Receptors, Urokinase Plasminogen Activator; D012711:Serum Albumin, Radio-Iodinated; D005340:Fibrinogen; D010958:Plasminogen; D010959:Tissue Plasminogen Activator; D005341:Fibrinolysin