Association of thrombin, plasmin, thrombin-antithrombin III complex and plasmin-antithrombin III complex with isolated hepatocytes.
The interaction of thrombin, plasmin or their antithrombin III complexes with isolated mouse hepatocytes was studied. Plasmin bound to hepatocytes in a concentration-dependent manner with an apparent Kd of 6.4.10(-8) M, attaining equilibrium within 10 min, and the interaction was inhibited by 6-amino-n-hexanoic acid. Plasmin treated with diisopropylfluorophosphate (DFP) bound to the cells in similar way as the untreated form of the enzyme. Thrombin bound also to hepatocytes, in a concentration-dependent manner, with a Kd of 5.4.10(-8) M reaching a steady state after 180 min. Thrombin inactivated with DFP, however, was inhibited in its binding to these cells. These data suggest that, whereas the kringle domains of plasmin are responsible for the enzyme-cell interaction, the active center of thrombin may be involved in the binding of this enzyme to hepatocytes. Plasmin-antithrombin III and thrombin-antithrombin III complexes were also associated with hepatocytes in a time-dependent manner, reaching a plateau after 180 min, and the two complexes competed in the interaction. While the interaction of active proteinases plasmin or thrombin with hepatocytes did not result in their internalization, the antithrombin III complexes were taken up by the cells, and thrombin-antithrombin III complex was degraded. These results indicate that hepatocytes may participate in the elimination of proteinase-antithrombin III complexes from the plasma, while the association of plasmin and thrombin with hepatocytes could imply distinct biological importance.
Spolarics
Z
Z
First Institute of Biochemistry, Semmelweis University Medical School, Budapest, Hungary.
Kalapos
M P
MP
Léránt
I
I
Garzó
T
T
Antoni
F
F
Mandl
J
J
Machovich
R
R
D000818:Animals; D000990:Antithrombin III / Q000378:metabolism*; D001667:Binding, Competitive; D002469:Cell Separation; D005341:Fibrinolysin / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D008099:Liver / Q000166:cytology / Q000378:metabolism*; D046911:Macromolecular Substances; D051379:Mice; D010447:Peptide Hydrolases / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000032:analysis / Q000187:drug effects; D018179:Receptors, Thrombin; D013917:Thrombin / Q000378:metabolism*; D014357:Trypsin / Q000494:pharmacology
D046911:Macromolecular Substances; D011956:Receptors, Cell Surface; D018179:Receptors, Thrombin; D000990:Antithrombin III; D010447:Peptide Hydrolases; D014357:Trypsin; D013917:Thrombin; D005341:Fibrinolysin