Differential binding properties of human pregnancy zone protein- and alpha2-macroglobulin-proteinase complexes to low-density lipoprotein receptor-related protein.
Human pregnancy zone protein (PZP) is a major pregnancy-associated plasma protein strongly related to alpha2-macroglobulin (alpha2-M). Both alpha-macroglobulins (alpha-Ms) covalently bind proteinases, which is accompanied by the exposure of carboxy terminal receptor recognition domains important for the rapid clearance from the circulation and tissues. It is accepted that the molecule responsible for the clearance of alpha2-M- and PZP-proteinase complexes is the low-density lipoprotein receptor-related protein (LRP). Although both alpha-M-proteinase complexes bind to the same receptor, differences in the binding properties have been reported. In addition, although it is known that the binding of alpha2-M-proteinase complexes to LRP can be blocked by Ni2+, the effect on PZP-proteinase has never been examined. In order to investigate differences in the binding properties of both alpha-Ms to the receptor, we purified LRP from human placenta by affinity chromatography and then analyzed the specificity and affinity of binding of alpha2-M- and PZP-proteinase complexes to the receptor by enzyme immunoassay. Our results clearly established that although both alpha-M-proteinase complexes specifically bind to LRP, PZP-chymotrypsin complexes bind to the receptor with lesser apparent affinity (Kd approximately equal 320 nM) than alpha2-M-chymotrypsin complexes (Kd approximately equal 40 nM). We also demonstrated that Ni2+ blocks the binding of alpha2-M-chymotrypsin complexes, but not PZP-chymotrypsin complexes, to LRP. These data suggest that the binding to LRP involves conformational differences between both alpha-Ms in a region immediately upstream of the carboxy terminal receptor recognition domain. The possibility that PZP-proteinase complexes interact with other receptors not available to alpha2-M-proteinase complexes could be considered.
Chiabrando
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GA
Departamento de Bioquímica Clínica, Universidad Nacional de Córdoba, Córdoba, 5000, Argentina. gustavo@bioclin.fcq.unc.edu.ar
Vides
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Sánchez
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D001667:Binding, Competitive; D010450:Endopeptidases / Q000378:metabolism*; D006801:Humans; D026502:LDL-Receptor Related Proteins / Q000378:metabolism*; D008024:Ligands; D009532:Nickel / Q000494:pharmacology; D011257:Pregnancy Proteins / Q000378:metabolism*; D000511:alpha-Macroglobulins / Q000378:metabolism*
D026502:LDL-Receptor Related Proteins; D008024:Ligands; D011257:Pregnancy Proteins; D000511:alpha-Macroglobulins; D009532:Nickel; D010450:Endopeptidases