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  Interaction of plasminogen activator inhibitor type-1 (PAI-1) with vitronectin - Characterization of different PAI-1 mutants

De Prada, N. A., Schroeck, F., Sinner, E. K., Mühlenweg, B., Twellmeyer, J., Sperl, S., et al. (2002). Interaction of plasminogen activator inhibitor type-1 (PAI-1) with vitronectin - Characterization of different PAI-1 mutants. European Journal of Biochemistry, 269(1), 184-192.

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Genre: Journal Article
Alternative Title : Eur. J. Biochem.

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 Creators:
De Prada, N. A., Author
Schroeck, F., Author
Sinner, E. K.1, Author           
Mühlenweg, B., Author
Twellmeyer, J.2, Author           
Sperl, S.2, Author           
Wilhelm, O. G., Author
Schmitt, M., Author
Magdolen, V., Author
Affiliations:
1Oesterhelt, Dieter / Membrane Biochemistry, Max Planck Institute of Biochemistry, Max Planck Society, ou_1565164              
2External Organizations, ou_persistent22              

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Free keywords: plasminogen activator inhibitor type-1; vitronectin; heparin; mutational analysis; surface plasmon resonance
 Abstract: The serpin plasminogen activator inhibitor type 1 (PAI-1) plays an important role in physiological processes such as thrombolysis and fibrinolysis, as well as pathophysiological processes such as thrombosis, tumor invasion and metastasis. In addition to inhibiting serine proteases, mainly tissue-type (tPA) and urokinase-type (uPA) plasminogen activators, PAI-1 interacts with different components of the extracellular matrix, i.e. fibrin, heparin (Hep) and vitronectin (Vn). PAI-1 binding to Vn facilitates migration and invasion of tumor cells. The most important determinants of the Vn-binding site of PAI-1 appear to reside between amino acids 110-147, which includes alpha helix E (hE, amino acids 109-118). Ten different PAI-1 variants (mostly harboring modifications in hE) as well as wild-type PAI-1, the previously described PAI-1 Mutant Q123K, and another serpin, PAI-2, were recombinantly produced in Escherichia coli containing a His(6) tag and purified by affinity chromatography. As shown in microtiter plate-based binding assays, surface plasmon resonance and thrombin inhibition experiments, all of the newly generated mutants which retained inhibitory activity against uPA still bound to Vn. Mutant A 114-118, in which all amino-acids at positions 114-118 of PAI-1 were exchanged for alanine, displayed a reduced affinity to Vn as compared to wildtype PAI-1. Mutants lacking inhibitory activity towards uPA did not bind to Vn. Q123K, which inhibits uPA but does not bind to Vn, served as a control. In contrast to other active PAI-1 mutants, the inhibitory properties of A 114-118 towards thrombin as well as uPA were significantly reduced in the presence of Hep. Our results demonstrate that the wild-type sequence of die region around hE in PAI-1 is not a prerequisite for binding to Vn.

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Language(s): eng - English
 Dates: 2002-01
 Publication Status: Published in print
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: eDoc: 41528
ISI: 000173398100021
 Degree: -

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Title: European Journal of Biochemistry
  Alternative Title : Eur. J. Biochem.
Source Genre: Journal
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Publ. Info: -
Pages: - Volume / Issue: 269 (1) Sequence Number: - Start / End Page: 184 - 192 Identifier: ISSN: 0014-2956