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| 蛋白免疫共沉淀筛选肾癌A498细胞CXCR4核定位结合蛋白 |
| 王志向1,2△,蔡晨3△,王林辉1,刘冰1,杨庆1,孙颖浩1* |
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(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*通信作者) |
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| 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。 |
| 关键词: 肾肿瘤 NR1D2 c-src HSPA8 CXCR4核定位 免疫共沉淀 |
| DOI：10.3724/SP.J.1008.2013.0068 |
| 投稿时间：2012-10-16修订日期：2012-11-13 |
| 基金项目:国家自然科学基金(81272817/H1619). |
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| Co-immunoprecipitation in screening proteins interacting with CXCR4 during nuclear localization in renal cell carcinoma A498 cells |
| WANG Zhi-xiang1,2△,CAI Chen3△,WANG Lin-hui1,LIU Bing1,YANG Qing1,SUN Ying-hao1* |
(1. Department of Urology, Changhai Hospital, Second Military Medical University, Shanghai 200433, China
2. Department of Urology, No.188 Hospital of PLA, Chaozhou 521000, Guangdong, China
3. Department of Special Diagnosis, Changhai Hospital, Second Military Medical University, Shanghai 200433, China
△Co-first authors.
*Corresponding author.) |
| Abstract: |
| 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. |
| Key words: kidney neoplasms NR1D2 c-src HSPA8 CXCR4 nuclear localization co-immunoprecipitation |
