Biosynthesis and processing of cathepsins B,H and L in carp macrophages

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Cathepsins B,H,and L were purified from carp hepato-pancreas or muscle to prepare their antibodies.15 resudures of N-terminal amino acid sequences of cathepsin L were determined. Polyclonal antibodies of cathepsins B and L,and monoclonal antibody of cathepsin L were prepared by using purified enzymes.We established the method of preparation of macrophage monolayr from carp peritoneal exudate cells.The intracellular processing and release of cathepsins B and L by carp peritoneal macrophages were investigated by pulse-chase experiments. Newry synthesized procathepsin B (39 kDa) after 15 min labeling was processed to the mature, single-chain enzymc (29 kDa) within 1 h. However two-chain form of cathepsin B was not detected during 24 h.Though processing of cathepsin L was not clarified, cathepsi L was synthesized as procathepsin L of 39 kDa.