Purification and characterization of cathepsin H from hepatopancreas of carp Cyprinus carpio.
1. Cathepsin H was purified about 5400-fold from hepatopancreas of carp (Cyprinus carpio) by the method involving ammonium sulfate fractionation, and chromatography on S-Sepharose, DEAE-Sephacel, Ultrogel AcA54, Concanavalin A-Sepharose 4B and GPC on Protein-Pak 125. 2. The purified cathepsin H gave a single protein band on analytical-PAGE, but migrated as two bands of 27,000 and 23,000 mol. wt on SDS-PAGE. 3. Cathepsin H had a pH and temperature optimum of 6.5 and 45 degrees C using Arg-MCA as a substrate, respectively, and was activated by sulfhydryl compounds and inhibited by cysteine protease inhibitors and metal compounds having high reactivities at cysteine residue. 4. The carp hepatopancreas cathepsin H immunoreacted with the monospecific antibody against rat liver cathepsin H, and did not react with the antibodies against carp hepatopancreas cathepsins B and L by the method of immunoelectrophoretic blotting.
Aranishi
F
F
Laboratory of Marine Biological Chemistry, Faculty of Fisheries, Nagasaki University, Japan.
Hara
K
K
Ishihara
T
T
D000818:Animals; D002347:Carps / Q000378:metabolism*; D056655:Cathepsin H; D002403:Cathepsins / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification*; D002614:Chelating Agents / Q000494:pharmacology; D003546:Cysteine Endopeptidases; D004795:Enzyme Stability; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D007120:Immunochemistry; D008099:Liver / Q000201:enzymology; D008970:Molecular Weight; D010179:Pancreas / Q000201:enzymology; D013438:Sulfhydryl Compounds / Q000494:pharmacology
D002614:Chelating Agents; D013438:Sulfhydryl Compounds; D002403:Cathepsins; D003546:Cysteine Endopeptidases; D056655:Cathepsin H; C534128:Ctsh protein, rat