甲醛和苯并(a)芘单独染毒条件下诱导型热休克蛋白70的交互作用蛋白分析
首发时间:2013-12-11
摘要: 目的 分析鉴定人支气管上皮细胞(HBE)在甲醛和苯并(a)芘(BaP)单独染毒时,诱导型热休克蛋白70(Hsp70)的所有交互作用蛋白,为分析Hsp70在甲醛和苯并(a)芘单独染毒时所发挥的生物学作用的异同提供依据 方法 用3.3mM的甲醛和16μM 的BaP单独染毒人支气管上皮细胞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 甲醛染毒时的Hsp70交互作用蛋白经过鉴定共有41种,根据功能分为13类;BaP染毒阶段时的Hsp70交互作用蛋白经过鉴定总共有55种,根据功能分为13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。其中ANXA2和Glutathione S-transferase P在甲醛染毒和BaP染毒时均和Hsp70有交互作用。 结论 在甲醛和BaP单独染毒时,Hsp70可能通过和特定蛋白的交互作用而在加速毒物代谢解毒、减小损伤、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 separately
Abstract: In this paper, we separated and analyzed all the Hsp70-interacting proteins during exposure to formaldehyde and Benzo[a]pyrene (BaP) respectively, in order to compare the difference of biological function of Hsp70. We exposed HBE to 3.3mM formaldehyde or 16μM BaP for 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 exposure to BaP, there were 55 kinds of proteins and were divided into 13 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 and BaP through interacting with these proteins.
Keywords: Occupational health Hsp70 Co-immunoprecipitation Mass spectrum
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甲醛和苯并(a)芘单独染毒条件下诱导型热休克蛋白70的交互作用蛋白分析
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