Abstract
Vitronectin (VN) and plasminogen activator inhibitor-1 (PAI-1) have important functional interactions: VN stabilises the protease inhibitory activity of PAI-1 and PAI-1 inhibits binding of adhesion receptors to VN. Having previously mapped the PAI-1 binding area for VN, we have now constructed a PAI-1 variant, R103A-M112A-Q125A, without measurable affinity to VN, but with full protease inhibitory activity and endocytosis receptor binding. As a tool for evaluating the physiological and pathophysiological functions of the PAI-1-VN interaction, our new variant is far superior to the previously widely used PAI-1 variant Q125K, which we have found possesses an only about 10-fold reduced affinity to VN.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Substitution
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Anilino Naphthalenesulfonates / chemistry
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Anilino Naphthalenesulfonates / metabolism
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Antibodies, Monoclonal / genetics
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Antibodies, Monoclonal / pharmacology
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Binding, Competitive
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Cell Line
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Escherichia coli / metabolism
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Genetic Variation
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Half-Life
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Humans
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Inhibitory Concentration 50
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Low Density Lipoprotein Receptor-Related Protein-1 / metabolism
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Plasminogen Activator Inhibitor 1 / chemistry
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Plasminogen Activator Inhibitor 1 / genetics*
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Plasminogen Activator Inhibitor 1 / metabolism*
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Plasminogen Activator Inhibitor 1 / pharmacology
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Radioligand Assay
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Recombinant Proteins / chemistry
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Recombinant Proteins / genetics
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Recombinant Proteins / metabolism
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Recombinant Proteins / pharmacology
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Surface Plasmon Resonance
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U937 Cells
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Urokinase-Type Plasminogen Activator / chemistry
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Urokinase-Type Plasminogen Activator / metabolism
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Vitronectin / metabolism*
Substances
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Anilino Naphthalenesulfonates
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Antibodies, Monoclonal
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Low Density Lipoprotein Receptor-Related Protein-1
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Plasminogen Activator Inhibitor 1
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Recombinant Proteins
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Vitronectin
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Urokinase-Type Plasminogen Activator