Lysosomal degradation of alpha 2u-globulin and alpha 2u-globulin-xenobiotic conjugates.
A diverse group of chemicals cause a male rat-specific nephrotoxicity in which alpha 2u-globulin accumulates in renal lysosomes. It has been suggested that these chemicals bind to the protein and decrease its degradation by lysosomal proteinases. To test this hypothesis, the lysosomal degradation of native alpha 2u-globulin and that to which d-limonene, d-limonene-1,2-oxide, isophorone, 1,4-dichlorobenzene, and 2,5-dichlorophenol were bound was studied. alpha 2u-Globulin was purified from male rat urine, and male rat renal cortical lysosomes, isolated by differential centrifugation, served as the proteolytic enzyme source. Pepstatin, an inhibitor of aspartic acid proteinases, and leupeptin, an inhibitor of cysteine proteinases, reduced alpha 2u-globulin degradation to 28 +/- 8 and 17 +/- 5% of control, respectively, whereas addition of both inhibitors decreased alpha 2u-globulin degradation to 8 +/- 1% of control values. These results indicate that both classes of endopeptidases are important in the degradation of alpha 2u-globulin. Under the incubation conditions used, 30% of native alpha 2u-globulin was degraded in a 4-hr period. Conjugates of the protein were made for in vitro binding experiments. Binding of d-limonene and 1,4-dichlorobenzene to alpha 2u-globulin did not alter the degradation of the protein, whereas binding of d-limonene-1,2-oxide, 2.5-dichlorophenol, and isophorone decreased alpha 2u-globulin degradation by 33%. These results indicate that not all chemicals which have been shown to bind in vivo to alpha 2u-globulin alter the in vitro lysosomal degradation of the protein. However, in all cases, one metabolite of each hyaline droplet inducer did alter degradation of alpha 2u-globulin, suggesting that a decrease in lysosomal degradation is involved in the accumulation of this protein in male rat kidney lysosomes.
Miami Valley Laboratories, Procter and Gamble Co., Cincinnati, Ohio 45239-8707.
L D
LD
Lehman-McKeeman
M I
MI
Rivera-Torres
D
D
Caudill
D000510:Alpha-Globulins / Q000378:metabolism* / Q000494:pharmacology*; D000818:Animals; D016282:Aspartic Acid Endopeptidases; D002851:Chromatography, High Pressure Liquid; D000081005:Cyclohexane Monoterpenes; D003546:Cysteine Endopeptidases / Q000378:metabolism; D015853:Cysteine Proteinase Inhibitors; D004226:Dithioerythritol / Q000494:pharmacology; D010450:Endopeptidases / Q000378:metabolism; D007687:Kidney Tubules, Proximal / Q000187:drug effects; D008247:Lysosomes / Q000378:metabolism*; D008297:Male; D039821:Monoterpenes; D011485:Protein Binding; D051381:Rats; D011919:Rats, Inbred Strains; D013729:Terpenes / Q000494:pharmacology; D015262:Xenobiotics / Q000378:metabolism* / Q000494:pharmacology*
D000510:Alpha-Globulins; D000081005:Cyclohexane Monoterpenes; D015853:Cysteine Proteinase Inhibitors; D039821:Monoterpenes; D013729:Terpenes; D015262:Xenobiotics; C013803:alpha 2u globulin; C028940:limonene-1,2-epoxide; D004226:Dithioerythritol; D010450:Endopeptidases; D003546:Cysteine Endopeptidases; D016282:Aspartic Acid Endopeptidases