Construction of a plasminogen activator inhibitor-1 variant without measurable affinity to vitronectin but otherwise normal.
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.
Jensen
Jan K
JK
Laboratory of Cellular Protein Science, Department of Molecular Biology, Aarhus University, Gustav Wieds Vej 10C, 8000 C, Aarhus, Denmark. jkj@mb.au.dk
Durand
Michelle K V
MK
Skeldal
Sune
S
Dupont
Daniel M
DM
Bødker
Julie S
JS
Wind
Troels
T
Andreasen
Peter A
PA
D019943:Amino Acid Substitution; D000817:Anilino Naphthalenesulfonates / Q000737:chemistry / Q000378:metabolism; D000911:Antibodies, Monoclonal / Q000235:genetics / Q000494:pharmacology; D001667:Binding, Competitive; D002460:Cell Line; D004926:Escherichia coli / Q000378:metabolism; D014644:Genetic Variation; D006207:Half-Life; D006801:Humans; D020128:Inhibitory Concentration 50; D026503:Low Density Lipoprotein Receptor-Related Protein-1 / Q000378:metabolism; D017395:Plasminogen Activator Inhibitor 1 / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism* / Q000494:pharmacology; D011869:Radioligand Assay; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D020349:Surface Plasmon Resonance; D020298:U937 Cells; D014568:Urokinase-Type Plasminogen Activator / Q000737:chemistry / Q000378:metabolism; D019096:Vitronectin / Q000378:metabolism*
D000817:Anilino Naphthalenesulfonates; D000911:Antibodies, Monoclonal; D026503:Low Density Lipoprotein Receptor-Related Protein-1; D017395:Plasminogen Activator Inhibitor 1; D011994:Recombinant Proteins; D019096:Vitronectin; D014568:Urokinase-Type Plasminogen Activator