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

鲤鱼巨噬细胞组织蛋白酶B、H和L的生物合成和加工

• 批准号: 05660234
• 负责人: HARA Kenji
• 金额: $ 1.15万
• 依托单位: Nagasaki University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05660234/
• 关键词: Fish Cathepsins; Biosynthesis; Processing; Macrophage; Pulse-chase experiment

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.

从鲤鱼肝胰和肌肉中分离纯化组织蛋白酶B、H和L，制备其抗体。用纯化的酶制备了组织蛋白酶B和L的多克隆抗体和单克隆抗体，建立了鲤鱼腹腔渗出液巨噬细胞单层的制备方法，采用脉冲追踪实验研究了鲤鱼腹腔巨噬细胞对组织蛋白酶B和L的胞内加工和释放。Newry合成的组织蛋白原B（39 kDa）经15 min标记后，在1 h内被加工成成熟的单链酶（29 kDa）。然而，在24 h内未检测到双链形式的组织蛋白酶B。尽管组织蛋白酶L的加工尚不清楚，但组织蛋白酶L被合成为39 kDa的组织蛋白酶原L。

项目成果

原研治,槌本六秀,橘勝康,長富潔,古場久代,石原忠: "コイ腹腔マクロファージの調製" 長崎大学水産学部研究報告. 第74,75合併号. 51-57 (1993)

原健二、土本六秀、立花克泰、永富清、古叶久代、石原正：“鲤鱼腹腔巨噬细胞的制备”，长崎大学水产学部研究报告合订第74号和第75号。51-57（1993年）。