Involvement of a cystatin-alpha-sensitive cysteine proteinase in the degradation of native L-lactate dehydrogenase and serum albumin by rat liver or kidney lysosomes.
A lysosomal cysteine proteinase plays a critical role in the lysosomal degradation of native L-lactate dehydrogenase. The effects of a recombinant cystatin alpha and other cysteine proteinase inhibitors on the degradation of unlabeled native L-lactate dehydrogenase by total lysosomal enzymes were examined in vitro. L-Lactate dehydrogenase was inactivated, its 35-kDa subunit disappeared and the final amino acid degradation products were produced during an incubation with disrupted lysosomes in vitro. These processes were all markedly suppressed by a small amount of cystatin alpha without inhibiting the activities of the known lysosomal cysteine proteinases, cathepsins B, H, L, and J. These results suggest that a cysteine proteinases, which is highly sensitive to cystatin alpha and distinct from other known lysosomal cysteine proteinases, is involved in the lysosomal degradation of native L-lactate dehydrogenase. An L-lactate-dehydrogenase-inactivating enzyme in the extracts of lysosomes was partially purified. It was separated from cathepsin J using a Sephacryl S-200 gel-filtration column and was further separated from cathepsin H by DEAE-Sephadex A-50 anion-exchange column chromatography. The inactivation and degradation of L-lactate dehydrogenase by this partially purified enzyme were all markedly suppressed by a low level of cystatin alpha without inhibiting the activities of both cathepsins B and L. The degradation of rat serum albumin by the partially purified enzyme was also inhibited by the same concentration of cystatin alpha. It is concluded that a cystatin-alpha-sensitive cysteine proteinase, other than cathepsins B, H, L and J, is present in lysosomes and functions in the lysosomal degradation of at least native L-lactate dehydrogenase and serum albumin.[1]References
- Involvement of a cystatin-alpha-sensitive cysteine proteinase in the degradation of native L-lactate dehydrogenase and serum albumin by rat liver or kidney lysosomes. Ohshita, T., Kido, H. Eur. J. Biochem. (1994) [Pubmed]
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