Characterization and distribution of albumin binding protein in normal rat kidney.

The mechanism by which proteins that pass through the glomerular basal lamina are taken up by proximal tubule cells is incompletely characterized. Past work has identified the kinetics of albumin binding to renal brush-border membrane. We have now purified and characterized albumin binding protein (ABP) and shown its distribution in renal proximal tubular cells. ABP was purified from rat renal proximal tubular cell brush-border membrane by affinity chromatography with rat serum albumin-Sepharose. The resulting ABP had two apparent molecular masses (55 and 31 kDa) by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Antibodies to ABP were raised in rabbits and checked by immunoassay and immunoblotting. Light-microscopic immunohistochemistry showed ABP all along the proximal tubule in the pars convoluta and pars recta. Electron-microscopic immunohistochemistry showed labeling on microvilli and in apical endocytic vacuoles, dense apical tubules, and lysosomes. These results indicate that ABP is involved in proximal tubule endocytosis.

Transport of serum transthyretin into chicken oocytes. A receptor-mediated mechanism.

Transthyretin (TTR) is involved in the transport of thyroid hormones and, due to its interaction with serum retinol-binding protein, also of vitamin A. The importance of both ligands in vertebrate embryonic development has prompted us to investigate the molecular details of TTR transport function in a powerful germ cell system, the rapidly growing chicken oocytes. Yolk TTR is derived from the circulatory system, since biotinylated TTR was recovered by immunoaffinity chromatography of yolk obtained from a hen previously infused with in vitro biotinylated chicken serum proteins. In concordance with the intraoocytic localization in an endosomal compartment, ligand blotting and chemical cross-linking experiments revealed the presence of a approximately 115-kDa TTR-binding oocyte membrane protein. This putative TTR receptor was not detected in chicken ovarian granulosa cells or embryonic fibroblasts and was different from the previously described oocyte-specific receptor for two estrogen-induced chicken serum lipoproteins, vitellogenin and very low density lipoprotein (Barber, D. L., Sanders, E. J., Aebersold, R., and Schneider, W. J. (1991) J. Biol. Chem. 266, 18761-18770). Furthermore, in contrast to the serum levels of the yolk precursor lipoproteins, those of TTR were not significantly changed by estrogen; thus, TTR represents a newly defined, estrogen-independent class of yolk precursor proteins. These data strongly suggest that oocytic TTR is derived from the circulation, where it is a constitutive component, and deposited into yolk as a result of endocytosis mediated by a specific receptor.

Albumin-binding proteins on the surface of the Streptococcus milleri group and characterization of the albumin receptor of Streptococcus intermedius C5.

Members of the Streptococcus milleri group (SMG) that react with Lancefield group C antisera were shown to bind large amounts of albumin although there was no direct relation between these two properties as polyclonal antisera to Lancefield group C antigen did not prevent the binding of albumin. There was a specificity for albumin binding, with albumin from man, monkeys, cat, dog and mouse being bound to a greater degree than albumin from cow, horse, goat or rabbit. Gold-labelled albumin was shown to be located close to the surface of strains by transmission electron microscopy. A cell-surface protein of M(r) 24,000, which was liberated by lysozyme treatment of cells, was shown to be the cell-surface receptor on Streptococcus intermedius C5. The receptor was physically dissimilar from protein G, an albumin- and IgG-binding protein of 'large-colony' Lancefield group C and G streptococci.