免疫共沉淀联合质谱分析筛选克罗恩病中Intelectin-1相互作用蛋白的初步研究

摘要

Objective To screen the proteins interacted with Intelectin-1 in Crohn disease via comparing Crohn disease and normal tissues and investigate the function of Intelectin-1 in the development of Crohn disease.Methods The proteins interacting with Intelectin-1 were screened by co-immunoprecipitation and identified by MALDL-TOF/TOF-MS and finally confirmed by immunoprecipitation and Western blot analysis.Results Data of Western blot showed that Intelectin-1 was overexpressed in Crohn disease tissues compared with normal tissues. In addition, four novel proteins(ATPase,HSP90，TRAF3 and ZNF) interacted with Intelectin-1 were successfully verified by MALDL-TOF/TOF-MS. Conclusion Intelectin-1 involves in the development of Crohn disease via interacting with TRAF3, ATPase, HSP90 and ZNF.%目的 对比克罗恩病患者的病变与正常肠黏膜,筛选与肠凝集素-1(Intelectin-1)相互作用的差异蛋白,探讨Intelectin-1及其相互作用蛋白在克罗恩病发展中的作用.方法 利用免疫共沉淀技术筛选克罗恩病患者病变及正常组织中与Intelectin-1相互作用的蛋白;利用MALDI-TOF/TOF-MS质谱分析技术鉴定与Intelectin-1相互作用蛋白;并使用免疫共沉淀和Western blot分析技术验证所鉴定的Intelectin-1相互作用蛋白.结果 利用免疫共沉淀联合质谱分析技术成功筛选出4个与Intelectin-1相互作用差异蛋白,包括:肿瘤坏死因子受体相关因子3(TRAF3)、腺苷酸三磷酸酶(ATPase)、热休克蛋白90(HSP90)、锌指蛋白(ZNF).结论 Intelectin-1可能通过与TRAF3、ATPase、HSP90、ZNF蛋白相互作用,而影响克罗恩病的发展.