Characterization of the initial α-thrombin interaction with glycoprotein Ibα in relation to platelet activation

Mario Mazzucato, Luigi De Marco, Adriana Masotti, Paola Pradella, Wadie F. Bahou, Zaverio M. Ruggeri

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Abstract

We have evaluated the properties of α-thrombin interaction with platelets within 1 min from exposure to the agonist, a time frame during which most induced activation responses are initiated and completed. Binding at 37 °C was rapidly reversible and completely blocked by a monoclonal antibody, LJ-Ib10, previously shown to be directed against the α-thrombin interaction site on glycoprotein (GP) Ibα. By 2-5 min, however, binding was no longer fully reversible and was only partially inhibited by the anti-GP Ibα antibody. Results were similar at room temperature (22-25 °C), whereas the initial characteristics of α-thrombin interaction with platelets were preserved for at least 20 min at 4 °C. Equilibrium binding isotherms obtained at the latter temperature were compatible with a two-site model, but the component ascribed to GP Ibα, completely inhibited by LJ-Ib10, had 'moderate' affinity (k(d) on the order of 10-8 M) and relatively high capacity, rather than 'high' affinity (k(d) on the order of 10-10 M) and low capacity as currently thought. The parameters of α-thrombin binding to intact GP Ibα on platelets at 4 °C corresponded closely to those measured with isolated GP Ibα fragments regardless of temperature. Blocking the α- thrombin-GP Ibα interaction caused partial inhibition of ATP release and prevented the association with platelets of measurable proteolytic activity. These results support the concept that GP Ibα contributes to the thrombogenic potential of α-thrombin.

Original languageEnglish
Pages (from-to)1880-1887
Number of pages8
JournalJournal of Biological Chemistry
Volume273
Issue number4
DOIs
Publication statusPublished - Jan 23 1998

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Platelet Glycoprotein GPIb-IX Complex
Platelet Activation
Platelets
Thrombin
Chemical activation
Blood Platelets
Temperature
Isotherms
Glycoproteins
Adenosine Triphosphate
Monoclonal Antibodies
Association reactions
Antibodies

ASJC Scopus subject areas

  • Biochemistry

Cite this

Characterization of the initial α-thrombin interaction with glycoprotein Ibα in relation to platelet activation. / Mazzucato, Mario; De Marco, Luigi; Masotti, Adriana; Pradella, Paola; Bahou, Wadie F.; Ruggeri, Zaverio M.

In: Journal of Biological Chemistry, Vol. 273, No. 4, 23.01.1998, p. 1880-1887.

Research output: Contribution to journalArticle

Mazzucato, Mario ; De Marco, Luigi ; Masotti, Adriana ; Pradella, Paola ; Bahou, Wadie F. ; Ruggeri, Zaverio M. / Characterization of the initial α-thrombin interaction with glycoprotein Ibα in relation to platelet activation. In: Journal of Biological Chemistry. 1998 ; Vol. 273, No. 4. pp. 1880-1887.
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