Proteolytic machinery of glomerular epithelial cells against IgG

Accumulation of immune complexes in the subepithelial region of the glomerular basement membrane results in the lesion of membranous nephropathy. The inefficient handling of immune complexes by the glomerular epithelial cell was investigated by studying the mechanism of IgG proteolysis by the intracellular proteases of cultured epithelial cells. Radiolabelled IgG was incubated with extracts of cells and the digestion of IgG was monitored by SDS-PAGE analysis. Prolonged incubation of IgG with the cell extracts resulted in only partial degradation of the IgG. The enzyme responsible for the breakdown was determined to be the lysosomal cathepsin D based on the pH optimum and the presence of aspartate in the active site of the enzyme. SDS-PAGE analysis of the digestion fragments revealed that a large proportion of the incompletely degraded IgG was the (Fab)2 fragment, which was resistant to further proteolysis. This could be one of the possible explanations for the slow removal of IgG from the subepithelial location of the basement membrane.