Degradation of oxidized or oxidatively modified proteins is an essential part of the antioxidant defenses of cells. 4-Hydroxy-2-nonenal, a major reactive aldehyde formed by lipid peroxidation, causes many types of cellular damage. It has been reported that 4-hydroxy-2-nonenal-modified proteins are degraded by the ubiquitin-proteasome pathway or, in some cases, by the lysosomal pathway. However, our previous studies using U937 cells showed that 4-hydroxy-2-nonenal-modified glyceraldehyde-3-phosphate dehydrogenase is degraded by cathepsin G. In the present study, we isolated the 4-hydroxy-2-nonenal-modified glyceraldehyde-3-phosphate dehydrogenase-degrading enzyme from rat neutrophils to an active protein fraction of 28？kDa. Using the specific antibody, the 28？kDa protein was identified as cathepsin G. Moreover, the degradation activity was inhibited by cathepsin G inhibitors. These results suggest that cathepsin G plays a crucial role in the degradation of 4-hydroxy-2-nonenal-modified glyceraldehyde-3-phosphate dehydrogenase. 1. Introduction Low-to-moderate concentrations of reactive oxygen species affect a great number of physiological functions. However, when the reactive oxygen species concentration exceeds the antioxidative capacity of an organism, animal cells enter a state termed oxidative stress, in which the excess reactive oxygen species induces oxidative damage on cellular components. As a result, oxidative stress has been implicated in a large range of diseases, including cancer, diabetes, male infertility, autoimmune diseases, atherosclerosis, and cardiovascular disorders [1–3]. Exposure to oxidative stress, which occurs in the presence of reactive oxygen species and free radicals, causes many adverse events including modification of proteins and reactions with DNA [4]. Lipid peroxidation also occurs, and various reactive aldehydes, such as 2-alkenals, 4-hydroxy-2-alkenals, and ketoaldehydes, are generated [5]. 4-Hydroxy-2-nonenal (HNE) is a major reactive aldehyde formed by the peroxidation of ω-6 polyunsaturated fatty acids, such as linoleic acid and arachidonic acid [6], and is involved in various physiological and pathological reactions [5, 6]. HNE has a high stability and reactivity and is largely responsible for many types of cellular damage associated with oxidative stress. HNE covalently binds to cysteine, lysine, or histidine residues [5]. These adducts are detected in several disease lesions, including in the brains of patients with Alzheimer’s disease [7, 8] and in low-density lipoproteins of atherosclerotic lesions [9]. HNE
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