Purification and characterization of cathepsin S from hepatopancreas of carp Cyprinus carpio |
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Authors: | Henneke Pangkey Keniji Hara Katsuyasu Tachibana Min-Jie Cao Kiyoshi Osatomi and Tadashi Ishihara |
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Affiliation: | Graduate School of Marine Science and Engineering, Nagasaki University, Nagasaki, Nagasaki 852-8521, Japan |
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Abstract: | SUMMARY: Cathepsin S was purified from carp hepatopancreas to homogeneity up to 300-fold. The amino acid sequence of its NH2-terminus was determined to be V-P-D-A-M-D-W-Y-N-K-G-Y-V-T-D-V-K-N-Q. On the contrary, that of purified cathepsin L from carp hepatopancreas was to be V-P-N-S-L-D-W-R-E-K-G. Purified cathepsin S consisted of a single chain with 37 kDa estimated by sodium dodecylsulfate–polyacrylamide gel electrophoresis. The enzyme had strong hydrolytic activity toward Z-Phe-Arg-MCA with the pH optimum of 7.0, but this lacked the ability to hydrolyze most of the other MCA substrates. The optimum pH of cathepsin S for protein substrate (carp myosin heavy chain) was also to be pH 7.0. These properties of purified cathepsin S obviously differ from cathepsins B and L. The enzyme activity was totally inhibited by E-64, leupeptin, 5–5'-dithiobis (2-nitro-benzoic acid) and p -tosyl-lys chloromethylketone as well. |
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Keywords: | carp cathepsin L cathepsin S cysteine proteinase hepatopancreas |
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