Megalin/gp330 mediates uptake of albumin in renal proximal tubule

Serum albumin filtered in renal glomeruli is reabsorbed very efficiently in the proximal tubule by endocytosis. The present study was undertaken to determine whether megalin/gp330 binds and mediates endocytosis of albumin. Rat serum albumin (RSA) labeled with I-125 and colloidal gold particles labeled with bovine serum albumin (RSA) were microinfused into rat-surface proximal tubules in vivo, and tubular uptake was determined in the presence or absence of different substances known to interfere with ligand binding to megalin. Binding of I-125-BSA and I-125-RSA to purified megalin was also determined directly using Sepharose columns. The results revealed that the tubular uptake of I-125-labeled RSA was significantly inhibited by receptor-associated protein (RAP), which reduced the uptake by >50% and by cold RSA. The uptake of BSA gold by the proximal tubule was very intensive. BSA gold was found in small and large endocytic vacuoles, dense apical tubules, and in lysosomes. The uptake was reduced by RAP to 17%, by EDTA to 19%, by BSA to 16%, by megalin to 35%, by cytochrome c to 49%, and, together with gentamicin, there was virtually no uptake. Megalin-Sepharose columns bound I-125-labeled BSA as well as I-125-RSA, the binding was inhibited by RAP and EDTA, and analysis of the eluate revealed the bound tracer to be albumin. In conclusion, the present study demonstrates that megalin is a mediator of albumin reabsorption in renal proximal tubules.