Hypochlorous acid-modified low-density lipoprotein inactivates the lysosomal protease cathepsin B: protection by ascorbic and lipoic acids.
Unregulated uptake of oxidized LDL by the scavenger receptor(s) of macrophages is thought to be an early event in atherosclerotic lesion development. Accumulation of oxidized LDL within macrophages may result from resistance of the modified LDL to enzymatic hydrolysis or from direct inactivation of lysosomal enzymes by reactive LDL-associated moieties. Since HOCl-modified LDL has been detected in vivo, the effects of HOCI-modified LDL on the activities of the cysteine protease cathepsin B and the aspartyl protease cathepsin D were investigated. LDL (0.5 mg protein/ml), which had been exposed to HOCl (25-200 microM), caused rapid dose-dependent inactivation of cathepsin B, but not of cathepsin D. Exposure of LDL to HOCl results primarily in the formation of LDL-associated chloramines, and the model chloramine N(alpha)-acetyl-lysine chloramine also caused dose-dependent inactivation of cathepsin B. Incubation of HOCl-modified LDL with ascorbic and lipoic acids (25-200 microM) resulted in dose-dependent reduction of LDL-associated chloramines and concomitant protection against cathepsin B inactivation. Thus, the data indicate that HOCl-modified LDL inactivates cathepsin B by a chloramine-dependent mechanism, most likely via oxidation of the enzyme's critical cysteine residue. Furthermore, small molecule antioxidants, such as ascorbic and lipoic acids, may be able to inhibit this potentially pro-atherogenic process by scavenging LDL-associated chloramines.
Carr
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Linus Pauling Institute, Oregon State University, Corvallis 97331-6512, USA. anitra.carr@orst.edu
D000588:Amines / Q000378:metabolism; D001205:Ascorbic Acid / Q000378:metabolism* / Q000494:pharmacology; D002401:Cathepsin B / Q000378:metabolism*; D002402:Cathepsin D / Q000378:metabolism; D002700:Chloramines / Q000378:metabolism / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D010450:Endopeptidases / Q000378:metabolism*; D006801:Humans; D006997:Hypochlorous Acid / Q000494:pharmacology*; D007150:Immunohistochemistry; D008077:Lipoproteins, LDL / Q000378:metabolism*; D008247:Lysosomes / Q000201:enzymology; D008063:Thioctic Acid / Q000378:metabolism* / Q000494:pharmacology
D000588:Amines; D002700:Chloramines; D008077:Lipoproteins, LDL; D006997:Hypochlorous Acid; D008063:Thioctic Acid; D010450:Endopeptidases; D002401:Cathepsin B; D002402:Cathepsin D; D001205:Ascorbic Acid