Determination of the vitronectin binding site on plasminogen activator inhibitor 1 (PAI-1).
Vitronectin is the carrier protein of plasminogen activator inhibitor 1 (PAI-1). We used a well-characterized panel of anti-human PAI-1 monoclonal antibodies (MoAbs) to localize the vitronectin-binding site on PAI-1. By employing a direct vitronectin/PAI-1 binding assay and two vitronectin-dependent inhibition assays, we demonstrate that the anti-PAI-1 MoAbs CLB-5, CLB-10, CLB-2C8 and I1, directed against different epitopes in the region between amino acids 110 and 145, prevent the interaction of PAI-1 with vitronectin. We conclude that the region between amino acids 110 and 145 of PAI-1 harbours an important determinant for the interaction with vitronectin.
van Meijer
M
M
Department of Biochemistry (K1-159), Academic Medical Center, University of Amsterdam, The Netherlands.
Gebbink
R K
RK
Preissner
K T
KT
Pannekoek
H
H
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D001665:Binding Sites; D000939:Epitopes / Q000032:analysis; D016326:Extracellular Matrix Proteins / Q000378:metabolism; D006023:Glycoproteins / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D016297:Mutagenesis, Site-Directed; D017395:Plasminogen Activator Inhibitor 1 / Q000096:biosynthesis / Q000737:chemistry* / Q000378:metabolism*; D017354:Point Mutation; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism; D010959:Tissue Plasminogen Activator / Q000037:antagonists & inhibitors; D019096:Vitronectin
D000911:Antibodies, Monoclonal; D000939:Epitopes; D016326:Extracellular Matrix Proteins; D006023:Glycoproteins; D017395:Plasminogen Activator Inhibitor 1; D011994:Recombinant Proteins; D019096:Vitronectin; D010959:Tissue Plasminogen Activator