Characterization of the soluble form of the low density lipoprotein receptor-related protein (LRP).
We report characterization of the soluble form of the low density lipoprotein receptor-related protein (sLRP) which circulates in human plasma. Amino acid sequence analysis confirmed that sLRP isolated from human plasma contains the alpha-chain of LRP1. In addition, Western blot analysis identified a truncated beta-chain noncovalently associated with the purified alpha-chain. The molecular size (M(r) 55K) of the peptide portion of the truncated beta-chain indicates that the subunit comprises the extracellular portion of the beta-chain and terminates in a membrane-proximal region. We investigated the mechanism by which sLRP may be generated using the trophoblast cell line, BeWo, which releases sLRP in culture. Cell surface labeling experiments indicate that LRP is released from BeWo cells following expression at the cell surface. Incubation of BeWo cells in the presence of a metalloproteinase inhibitor, INH-3855-PI, results in a dose-dependent inhibition of LRP shedding. The metalloproteinase responsible for the shedding of LRP by BeWo cells is not up-regulated by phorbol ester and is not dependent on serine proteases, such as plasmin, for activity. The BeWo cell line is derived from a human gestational choriocarcinoma and preliminary studies suggest that LRP may be shed within the placenta during gestation. Increased levels of sLRP were detected in cord blood. In term placenta, LRP is expressed in the syncytium, which comprises the maternal-fetal interface. Increased levels of sLRP in cord blood may reflect cellular dysfunction and increased metalloproteinase activity at this important interface.
Quinn
K A
KA
Centre for Thrombosis and Vascular Research, School of Pathology, Kensington, New South Wales, 2052, Australia. k.quinn@unsw.edu.au
Pye
V J
VJ
Dai
Y P
YP
Chesterman
C N
CN
Owensby
D A
DA
D000595:Amino Acid Sequence; D019921:Biotinylation; D002822:Choriocarcinoma / Q000378:metabolism; D005260:Female; D006801:Humans; D026503:Low Density Lipoprotein Receptor-Related Protein-1; D008565:Membrane Proteins / Q000378:metabolism; D008666:Metalloendopeptidases / Q000037:antagonists & inhibitors; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry; D010920:Placenta / Q000502:physiology; D011110:Polymorphism, Genetic; D011233:Precipitin Tests; D011247:Pregnancy; D011485:Protein Binding; D011971:Receptors, Immunologic / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D017421:Sequence Analysis; D012995:Solubility; D010959:Tissue Plasminogen Activator / Q000378:metabolism; D014327:Trophoblasts / Q000502:physiology; D014407:Tumor Cells, Cultured; D000511:alpha-Macroglobulins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
D026503:Low Density Lipoprotein Receptor-Related Protein-1; D008565:Membrane Proteins; D010446:Peptide Fragments; D011971:Receptors, Immunologic; D000511:alpha-Macroglobulins; D010959:Tissue Plasminogen Activator; D008666:Metalloendopeptidases