Albondin-mediated capillary permeability to albumin. Differential role of receptors in endothelial transcytosis and endocytosis of native and modified albumins.

Abstract

Specific albumin binding to the surface of endothelium initiates its transcytosis across continuous endothelium via noncoated plasmalemmal vesicles. Past work has identified several putative albumin-binding proteins (SPARC, gp60, gp30, and gp18). In this study, we examined the specific role of these proteins in the binding of bovine serum albumin (BSA) to endothelium. The 60-kDa albumin-binding protein (gp60, now called albondin) was purified from cultured rat microvascular endothelial cells for antibody production. Anti-albondin antibodies (alpha gp60) specifically recognized albondin expressed by rat, bovine, and human endothelial cells (EC). alpha gp60 and unlabeled native BSA inhibited 125I-BSA binding to confluent EC monolayers and to both albondin and SPARC extracts immobilized on filters. Modification of BSA by maleic anhydride treatment (Mal-BSA) or by surface adsorption to colloidal gold particles (A-Au) renders the ligand specific for gp30 and gp18 while eliminating its ability to inhibit 125I-BSA binding to EC and to both albondin and SPARC extracts. Mal-BSA and A-Au interacted with EC via distinct binding sites not recognizing native BSA. EC internalization of 125I-BSA is inhibited by unlabeled BSA and alpha gp60 IgG but not nonimmune IgG, Mal-BSA, or A-Au. Conversely, internalization of modified BSA is inhibited by unlabeled modified BSA but not BSA or alpha gp60 IgG. Specific 125I-BSA transendothelial transport in rat lungs perfused in situ and for EC monolayers in vitro is inhibited (> or = 90%) by unlabeled BSA and alpha gp60 IgG but not nonimmune IgG and Mal-BSA. No specific transport of 125I-Mal-BSA is detected across bovine lung microvascular endothelial cell monolayers, only paracellular and/or fluid-phase transport. Low temperatures reduce BSA transport by 5-fold and Mal-BSA by 2-fold. Interestingly, 3-fold more native BSA is transported than Mal-BSA at 37 degrees C, whereas at 8-10 degrees C they are nearly equal, suggesting facilitation of BSA transport. Cumulatively, it appears that gp30 and gp18 mediate the binding, endocytosis, and degradation of modified albumins, whereas albondin mediates native albumin binding which significantly enhances its transcytosis and capillary permeability.