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The interaction of human alpha-thrombin with mini-pig aortic endothelial cells was studied using 125I-labeled enzyme. Equilibrium between bound and free thrombin was attained within 1 min, and the Klotz-Hunston equations indicated two populations of binding sites. Approximately 30,000 sites/cell belonged to the high-affinity class with a Kd of about 3 x 10(-8) M. Modification of two lysine residues of thrombin with pyridoxal 5'-phosphate (PLP2-thrombin) destroyed the high-affinity binding and about three-fourths of the low-affinity bindings. When the lysine residue of thrombin involved in heparin binding was protected with heparin against chemical modification (PLP-thrombin), the modified enzyme remained similar to the native one with respect to cellular binding, with some loss of low-affinity binding only. Heparin, in a tenfold molar excess to enzyme, inhibited the binding of the native as well as the PLP-thrombin, whereas it did not influence the interaction between PLP2-thrombin and the cell. Since heparin might interfere with both the enzyme and the cell, the binding of heparin to endothelial cells was also examined. The results revealed that 3H-heparin also bound to cells. This binding was characterized by a Kd of 3 x 10(-7) M, approximately 10(6) sites/cell. Furthermore, thrombin bound to endothelial cells was released by antithrombin III. On the basis of these and other data in the literature, a model is proposed for the mechanism of the binding of thrombin to endothelial cells.
P T Bauer, and
R Machovich, and
P Arányi, and
K G Büki, and
E Csonka, and
I Horváth
May 1979,
The Journal of biological chemistry,
P T Bauer, and
R Machovich, and
P Arányi, and
K G Büki, and
E Csonka, and
I Horváth
November 1980,
The Journal of biological chemistry,
P T Bauer, and
R Machovich, and
P Arányi, and
K G Büki, and
E Csonka, and
I Horváth
January 1981,
Biochemistry,
P T Bauer, and
R Machovich, and
P Arányi, and
K G Büki, and
E Csonka, and
I Horváth
January 2002,
American journal of physiology. Endocrinology and metabolism,
P T Bauer, and
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K G Büki, and
E Csonka, and
I Horváth
April 1993,
International journal of cancer,
P T Bauer, and
R Machovich, and
P Arányi, and
K G Büki, and
E Csonka, and
I Horváth
August 1994,
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P T Bauer, and
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K G Büki, and
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I Horváth
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British journal of haematology,
P T Bauer, and
R Machovich, and
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K G Büki, and
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I Horváth
January 1992,
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P T Bauer, and
R Machovich, and
P Arányi, and
K G Büki, and
E Csonka, and
I Horváth
January 1981,
Annals of the New York Academy of Sciences,
P T Bauer, and
R Machovich, and
P Arányi, and
K G Büki, and
E Csonka, and
I Horváth
May 1981,
The Journal of biological chemistry,
