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作 者:应跃斌[1] 孙红颖[1] 丁丁[1] 李丹曦[1] 薛乔[1] 陈枢青[1]
机构地区:[1]浙江大学药学院药理毒理与生化药学研究所,浙江杭州310058
出 处:《浙江大学学报(医学版)》2009年第5期505-510,共6页Journal of Zhejiang University(Medical Sciences)
摘 要:目的:探索影响金黄色葡萄球菌肠毒素C2(SEC2)断裂的因素。方法:将纯化的肠毒素在不同条件下处理,观察环境因素对其断裂程度的影响。结果:重组SEC2断裂的位置可能位于其分子93位半胱氨酸与110位半胱氨酸之间。重组SEC2在37℃下、24 h内会完全产生断裂,在碱性条件下则加速其断裂的形成;H2O2会导致重组SEC2产生非特异性降解。当溶液中有β-巯基乙醇(2%)、苯甲基磺酰胺(5-10 mmol/L)、咪唑(1 mol/L)或大肠杆菌粗提物时,重组SEC2的断裂可被有效抑制。结论:肠毒素C2容易在特定位点发生断裂,可能与蛋白质本身结构有关。Objective: To investigate the limited digestion of recombinant staphylococcal enterotoxin C2 (SEC2 His) in different conditions. Methods: The purified recombinant SEC2 His was treated with different reagents and the cleavage of rSEC2 molecule was observed by SDS- PAGE. Results: The cleavage occurred in positions Cys93-Cys110 of the disulfide loop. Complete auto-cleavage of recombinant SEC2 was observed in solution at 37℃ within 24 hrs,and that was accelerated under alkaline conditions. The auto cleavage of the recombinant protein was inhibited in the presence of β-ME (2%),PMSF (5-10 mmol/L),imidazole (1 mol/L) or crude E. coli iysate. Non-specific degradation of recombinant SEC2 was promoted with the increasing of the concentration of H2O2. Conclusion: The recombinant SEC2-His is broken down in special site of protein,which may be associated with the protein structure.
关 键 词:肠毒素类 葡萄球菌 金黄色 超抗原 重组融合蛋白质类 生物降解 温度 氢离子浓度 蛋白酶抑制药
分 类 号:R378[医药卫生—病原生物学]
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