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Thromb Haemost 1988; 60(03): 419-427
DOI: 10.1055/s-0038-1646983
Original Article
Schattauer GmbH Stuttgart

Characterization of the Binding of Bovine Thrombin to Isolated Rat Hepatocytes

Britta Weyer
1   The Institute of Physiology, University of Aarhus, Aarhus, Denmark
,
Torben E Petersen
2   The Institute of Molecular Biology, University of Aarhus, Aarhus, Denmark
,
Ole Sonne
1   The Institute of Physiology, University of Aarhus, Aarhus, Denmark
› Institutsangaben
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Publikationsverlauf

Received 10. November 1987

Accepted after revision 21. Juli 1988

Publikationsdatum:
30. Juni 2018 (online)

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Summary

Isolated rat hepatocytes possess per cell 4,500 high-affinity binding sites for thrombin with a Kd of 30-40 pM, and 2.8 × 105 low-affinity sites with a Kd of 30 nM. These binding sites are highly specific for thrombin. Half-maximal binding of 125I-labelled thrombin is achieved after 3 min at 37¸ C and 7 min at 4¸ C. The reversibly bound fraction of the ligand dissociates according to a biexponential time course with the rate constants 1—2 × 10−2 s−1 and 3—4 × 10−4 s−1. Part of the tracer remains cell-associated even after prolonged incubation, but all cell-associated radioactivity migrates as intact thrombin upon sodium dodecyl sulphate polyacrylamide gel electrophoresis. The bound thrombin is minimally endocytosed as judged by the resistance to pH 3-treatment. Cell-associated radioactivity dissociated from the cells binds just aswell in a receptor assay as tracer incubated in a conditioned medium under the same conditions, indicating the absence of a quantitatively important receptor-mediated degradation ofthe ligand.

Keywords

Thrombin - Receptor - Thrombin - binding site - High - affinity - binding - Rat hepatocyte
 

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