蛋白免疫共沉淀筛选肾癌A498细胞CXCR4核定位结合蛋白  

Co-immunoprecipitation in screening proteins interacting with CXCR4 during nuclear localization in renal cell carcinoma A498 cells

作　　者：王志向[1,2] 蔡晨[3] 王林辉[1] 刘冰[1] 杨庆[1] 孙颖浩[1] 

机构地区：[1]第二军医大学长海医院泌尿外科,上海200433 [2]解放军188医院泌尿外科,潮州521000 [3]第二军医大学长海医院特需诊疗科,上海200433

出　　处：《第二军医大学学报》2013年第1期68-71,共4页Academic Journal of Second Military Medical University

基　　金：国家自然科学基金(81272817/H1619)~~

摘　　要：目的筛选肾癌A498细胞CXCR4核定位结合蛋白。方法采用蛋白免疫共沉淀实验寻找特异性条带,再通过蛋白质谱分析、生物信息学分析寻找可能与CXCR4结合的蛋白。结果 CXCR4抗体进行蛋白免疫共沉淀,发现有3条特异性条带。选取特异性条带行质谱分析提示可能的蛋白共有36个。将这36个蛋白与CXCR4进行生物信息学分析发现NR1D2、c-src及HSPA8有可能与CXCR4相互作用,参与CXCR4核定位。结论 NR1D2、c-src及HSPA8可能参与了A498细胞CXCR4核定位的过程。Objective To screen for the proteins interacting with CXCR4 during nuclear localization in renal cell carcinoma （RCC） A498 cells. Methods Specific band in co-immunoprecipitation （Co-IP） experiments was sent for mass spectrometry. With the results of Co-IP experiments and mass spectrometry, the proteins interacting with CXCR4 were determined by bioinformatics analyses. Results Three specific bands were found after Co-IP with anti-CXCR4 antibody, and the results of mass spectrometry of the three specific bands showed 36 proteins possibly interacting with CXCR4. Bioinformatics analyses showed that NR1D2, c-src and HSPA8 might interact with CXCR4 and participate in CXCR4 nuclear localization. Conclusion NR1D2, c-src and HSPA8 might have participated in CXCR4 nuclear localization in RCC A498 cells.

关 键 词：肾肿瘤 NR1D2 C-SRC HSPA8 CXCR4核定位 免疫共沉淀 

分 类 号：R737.11[医药卫生—肿瘤]