Functional interaction of plasminogen activator inhibitor type 1 (PAI-1) and heparin
| Autor: | Raymond Klein Gebbink, Hartmut J. Ehrlich, Hans Pannekoek, Klaus T. Preissner, Jaap Keijer |
|---|---|
| Rok vydání: | 1991 |
| Předmět: |
Protein Conformation
medicine.medical_treatment Molecular Sequence Data Biochemistry Plasminogen Activators Thrombin medicine Humans Amino Acid Sequence Vitronectin Cells Cultured Glycoproteins Urokinase Heparin cofactor II Protease Binding Sites biology Heparin Sepharose Antithrombin Affinity Labels Molecular biology Plasminogen Inactivators biology.protein Heparin Cofactor II Plasminogen activator medicine.drug |
| Zdroj: | Biochemistry. 30(4) |
| ISSN: | 0006-2960 |
| Popis: | Plasminogen activator inhibitor type 1 (PAI-1), the fast-acting inhibitor of tissue-type plasminogen activator (t-PA) and urokinase (u-Pa), is a member of the serpin superfamily of proteins. Both in plasma and in the growth substratum of cultured endothelial cells, PAI-1 is associated with its binding protein vitronectin, resulting in a stabilization of active PAI-1. Recently, it has been demonstrated that the PAI-1-binding site on vitronectin is adjacent to a heparin-binding site (Preissner et al., 1990). Furthermore, it can be deduced that the amino acid residues, proposed to mediate heparin binding in the serpins antithrombin III and heparin cofactor II, are conserved in PAI-1. Consequently, here we have investigated whether PAI-1 also interacts with heparin. At pH 7.4, PAI-1 quantitatively binds to heparin-Sepharose and can be eluted with increasing [NaCl]. Binding of PAI-1 to heparin-Sepharose can be efficiently competed with heparin in solution (IL 50 , 7 μM). In the presence of heparin, the protease specificity of PAI-1 toward thrombin is substantially increased. This is shown by (i) quenching of thrombin activity of PAI-1 in the presence of heparin and (ii) induction of the formation of SDS-stable complexes between thrombin and PAI-1 by heparin. In a dose response curve, both effects reached a maximum at approximately unit/mL and then diminished again upon further increasing the heparin concentration, strongly suggesting a template mechanism as an explanation for the observed effect. In contrast to vitronectin, heparin does not stabilize the action conformation of PAI-1. We propose that PAI-1, like antithrombin III, heparin cofactor II, and protease nexin 1, belongs to the group of heparin-dependent serpins. The binding of PAI-1 to heparin suggests that heparin may contribute to the localization of PAI-1 at particular sites, thus being involved in the regulation of plasminogen activation. Furthermore, we provide evidence that heparin has the potential |
| Databáze: | OpenAIRE |
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