Bovine aortic endothelial cells (BAEs) were used as a model system to study the nature and origin of protein(s) in the extracellular matrix that bind to type 1 plasminogen activator inhibitor (PAI-1). Matrix samples were fractionated by SDS-PAGE and analyzed by PAI-1 ligand binding and by immunoblotting using antibodies to vitronectin (Vn). PAI-1 bound primarily to two Vn-related polypeptides of Mr 63,000 and 57,000, and both of these partially degraded polypeptides were present in the culture serum. Radiolabeling experiments failed to detect significant Vn biosynthesis by BAEs (less than 0.03% of total), or by human umbilical vein endothelial cells and HT 1080 cells. The binding of PAI-1 to Vn was relatively specific since direct binding studies failed to demonstrate significant interactions between PAI-1 and other matrix proteins (e.g., fibronectin, type IV collagen, laminin, or matrigel). Kinetic studies indicate that PAI-1 rapidly accumulates in the matrix when BAEs are plated on Vn, appearing in the conditioned medium only after a significant lag period (1-2 h). However, no PAI-1 was detected in the matrix when the cells were plated on fibronectin-coated dishes, and there was no lag period for PAI-1 accumulation in the medium. These results indicate that PAI-1 binds specifically to serum-derived Vn in the matrix, and suggest that the composition of both the matrix and serum itself may influence the pericellular distribution of this important inhibitor.
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September 01 1990
Serum-derived vitronectin influences the pericellular distribution of type 1 plasminogen activator inhibitor.
D Seiffert,
D Seiffert
Research Institute of Scripps Clinic, La Jolla, California 92037.
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N N Wagner,
N N Wagner
Research Institute of Scripps Clinic, La Jolla, California 92037.
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D J Loskutoff
D J Loskutoff
Research Institute of Scripps Clinic, La Jolla, California 92037.
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D Seiffert
Research Institute of Scripps Clinic, La Jolla, California 92037.
N N Wagner
Research Institute of Scripps Clinic, La Jolla, California 92037.
D J Loskutoff
Research Institute of Scripps Clinic, La Jolla, California 92037.
Online ISSN: 1540-8140
Print ISSN: 0021-9525
J Cell Biol (1990) 111 (3): 1283–1291.
Citation
D Seiffert, N N Wagner, D J Loskutoff; Serum-derived vitronectin influences the pericellular distribution of type 1 plasminogen activator inhibitor.. J Cell Biol 1 September 1990; 111 (3): 1283–1291. doi: https://doi.org/10.1083/jcb.111.3.1283
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