Abstract
The PAI-1 binding site for VN was studied using two independent methods. PAI-1 was cleaved by Staph V8 protease, producing 8 fragments, only 2 of which bound to [125I]-VN. These fragments were predicted to overlap between residues 91-130. Since PAI-2 has structural homology to PAI-1, but does not bind to vitronectin, chimeras of PAI-1 and PAI-2 were constructed. Four chimeras, containing PAI-1 residues 1-70, 1-105, 1-114, and 1-167 were constructed and expressed in vitro. PAI-1, PAI-2, and all of the chimeras retained inhibitory activity for t-PA, but only the chimera containing PAI-1 residues 1-167 formed a complex with VN. Together, these results predict that the VN binding site of PAI-1 is between residues 115-130.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Amino Acid Sequence
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Base Sequence
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Binding Sites
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DNA, Complementary / genetics
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Humans
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Molecular Sequence Data
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Peptide Fragments / 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 2 / chemistry
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Plasminogen Activator Inhibitor 2 / genetics
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Plasminogen Activator Inhibitor 2 / metabolism
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Polymerase Chain Reaction
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Protein Binding
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Protein Structure, Tertiary*
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Recombinant Fusion Proteins / chemistry*
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Recombinant Fusion Proteins / metabolism
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Serine Endopeptidases / metabolism
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Vitronectin / metabolism*
Substances
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DNA, Complementary
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Peptide Fragments
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Plasminogen Activator Inhibitor 1
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Plasminogen Activator Inhibitor 2
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Recombinant Fusion Proteins
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Vitronectin
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Serine Endopeptidases
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glutamyl endopeptidase