Direct lysosomal uptake of α2-microglobulin contributes to chemically induced nephropathy

Abstract

An abnormal accumulation of alpha 2-microglobulin (alpha 2 mu) in kidney lysosomes of male rats has been described in the nephropathy resulting from exposure to a variety of chemicals. The increment in lysosomal levels of alpha 2 mu cannot be explained by a decrease in its proteolytic susceptibility. Because a portion of alpha 2 mu resides in the cytosol of kidney cells, we decided to analyze whether this cytosolic form also contributes to the abnormal lysosomal accumulation of alpha 2 mu after exposure to chemicals. Intact kidney lysosomes were isolated from untreated or 2,2,4-trimethylpentane (TMP) treated rats, and their ability to take up alpha 2 mu was compared. alpha 2 mu can be directly transported into isolated lysosomes in the presence of the heat shock cognate protein of 73 kDa (hsc73). alpha 2 mu specifically binds to a lysosomal membrane glycoprotein of 96 kDa, previously identified as the receptor for the hsc73-mediated lysosomal pathway of protein degradation. In rats exposed to TMP, the specific lysosomal transport of alpha 2 mu increases, as well as the ability of lysosomes to directly transport other substrates for this pathway. The increased lysosomal transport is mainly due to an increase in the levels of the receptor protein in the lysosomal membrane. The hsc73-mediated lysosomal pathway contributes to the normal degradation of alpha 2 mu in rat kidney and liver, and the activity of this pathway is increased after exposure to TMP. Our results suggest that the chemically induced accumulation of cytosolic alpha 2 mu in lysosomes is mediated by an increased rate of direct uptake into lysosomes.