甲醛染毒和染毒恢复阶段诱导型热休克蛋白70的交互作用蛋白分析
首发时间:2013-09-18
摘要:目的 分析鉴定人支气管上皮细胞(HBE)在甲醛染毒和染毒恢复时,诱导型热休克蛋白70(Hsp70)的所有交互作用蛋白,为阐明甲醛暴露条件下Hsp70的生物学作用提供依据 方法 用3.3mM的甲醛染毒人支气管上皮细胞3小时和染毒3小时后恢复3小时,分别收集、裂解细胞,然后运用免疫共沉淀技术分离蛋白,运用质谱技术鉴定蛋白,查询数据库和文献资料后根据功能对鉴定的蛋白进行分类 结果 甲醛染毒阶段的Hsp70交互作用蛋白经过鉴定共有41种,根据功能分为13类;甲醛染毒恢复阶段的Hsp70交互作用蛋白经过鉴定总共有55种,根据功能分为20类。其中既有具有正常生理功能的蛋白,如骨架蛋白、糖和能量代谢蛋白、细胞连接蛋白等,也有具有抵抗损伤促进修复功能的蛋白,如抵抗氧化性损伤蛋白、毒物代谢解毒蛋白、DNA修复蛋白等 结论 在甲醛染毒和染毒恢复阶段,Hsp70可能通过和特定蛋白的交互作用而在加速甲醛代谢解毒、减小甲醛导致的生物学损伤、促进变性蛋白和损伤DNA的修复方面发挥了一定的作用。
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An analysis of Hsp70-interacting proteins in human bronchial epithelium during exposure to formaldehyde and recovery period
Abstract:In this paper, we separated and analyzed all the Hsp70-interacting proteins during exposure to formaldehyde and recovery period, in order to elucidate the biological function of Hsp70. We exposed HBE to 3.3mM formaldehyde for 3h or recovered for another 3h, then collected and fragmentated cells, after that, separated and identified Hsp70-interacting proteins by co-immunoprecipitation and mass spectrum. During exposure to formaldehyde, there were 41 kinds of Hsp70-interacting proteins which were divided into 13 groups according to their function. And during the recovery period, there were 55 kinds of proteins and were divided into 20 groups. Some of Hsp70-interacting proteins have physiological functions such as cytoskeleton, glycometabolism and cell junction, some of them play a role in reducing damage or promoting repair such as detoxification of formaldehyde and DNA repair. These findings hint that Hsp70 may play a role in reducing damage or promoting repair resulted from formaldehyde through interacting with these proteins.
Keywords: Occupational health Hsp70 Co-immunoprecipitation Mass spectrum
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甲醛染毒和染毒恢复阶段诱导型热休克蛋白70的交互作用蛋白分析
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