Proteolytic degradation of muscllarr proteins of red sea bream by using immuno-electron mycroscope and immunoblot analysis

免疫电子显微镜和免疫印迹分析对红鲷肌肉蛋白的蛋白水解降解

• 批准号: 10660198
• 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 Scientific Research (C)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10660198/
• 关键词: Red sea bream Muscle; Autolysis; Protease; Myofibrillar Proteins; Protease Inhibitor; 筋原繊維タンパク質

We examined the degradation of post-mortem myofibrillar proteins (myosin heavy chain, α-actinin, β-connectin and troponin I) of red sea bream Pagrus major in relation with internal proteases by using specific protease inhibitors. These proteins were considerably hydrolyzed with internal proteinase(s) at 25℃ for 3 days. We confirmed that Cathepsin L which is cysteine protease in skeretal muscle was inhibited by E-64, specific cysteine protease inhibitor, injected to duct of Cuvie. Degradation of myosin HC and β-connectin were strongly inhibited by serine protease inhibitor (leupeptin and DFP). It was suggested that myosin HC is hydrolyzed by serine protease (may be MBP, myosin binding serine proteinase). Limited degradation of α-actinin was inhibited by cysteine protease inhibitor(E-64). The findings suggests that α-actinin is degraded by cathepsins B, L or S. Hydrolysis of tropomyosin and troponin I were mainly inhibited by serine protease inhibitors (leupeptin and DFP) and secondly inhibited by cysteine protease inhibitor(E-64). It is suggested that these proteins are hydrolyzed by MBP and the cathepsins.

采用特异性蛋白酶抑制剂研究真鲷（Pagrus major）死后肌原纤维蛋白（肌球蛋白重链、α-辅肌动蛋白、β-连接蛋白和肌钙蛋白I）的降解与体内蛋白酶的关系。这些蛋白质在25℃下用内部蛋白酶水解3天后可显著水解。我们证实，半胱氨酸蛋白酶特异性抑制剂E-64注射到Cuvie管中可抑制骨骼肌中的半胱氨酸蛋白酶组织蛋白酶L。丝氨酸蛋白酶抑制剂亮抑酶肽（leupeptin）和DFP能强烈抑制肌球蛋白HC和β-连接蛋白的降解。有人认为肌球蛋白HC被丝氨酸蛋白酶（可能是MBP，肌球蛋白结合丝氨酸蛋白酶）水解。半胱氨酸蛋白酶抑制剂（E-64）可抑制α-辅肌动蛋白的有限降解。结果表明，α-辅肌动蛋白可被组织蛋白酶B、L或S降解。丝氨酸蛋白酶抑制剂亮抑酶肽和DFP主要抑制原肌球蛋白和肌钙蛋白I的水解，其次是半胱氨酸蛋白酶抑制剂E-64。这表明这些蛋白质被MBP和组织蛋白酶水解。

项目成果

K.Hara, H.Pangkey, M.Kurihara, K.Tachibana, M.-J.Cao, K.Osatomi and T.Ishihara: "Proteolytic Degradation of Fish Myofibrillar Proteins by Internal Proteases"Proc.9th JSPS Joint Sem. Mar. Fish. Sci.. 299-309 (1999)

K.Hara、H.Pangkey、M.Kurihara、K.Tachibana、M.-J.Cao、K.Osatomi 和 T.Ishihara：“内部蛋白酶对鱼肌原纤维蛋白的蛋白水解降解”Proc.9th JSPS Joint Sem。

Kenji Hara et al: "Proteolytic Degradation of Fish Myofibrillar Proieins by Internal Proieases"Proc.9th JSPS Joint Sem.Mar.Fish.Sci.. 299-309 (1999)

Kenji Hara 等人：“内部蛋白酶对鱼肌原纤维蛋白的蛋白水解降解”Proc.9th JSPS Joint Sem.Mar.Fish.Sci. 299-309 (1999)

Kenji Hara et al.: "Proteolytic Degradation of Fish Myofibrillar Proteins by Internal Proteases"Proc. 9th JSPS Joint Sem. Mar. Fish. Sci.. 279-309 (1999)

Kenji Hara 等人：“内部蛋白酶对鱼肌原纤维蛋白的蛋白水解降解”Proc。