Screening of citrullinated proteins in ten tumor cell lines
10.3760/cma.j.issn.0253-3766.2015.03.002
- VernacularTitle:筛选肿瘤细胞中的瓜氨酸化蛋白
- Author:
Wei ZHANG
1
;
Lining MA
;
Yun SUI
;
Xiaotian CHANG
Author Information
1. 250001山东省济南市第二人民医院内科
- Keywords:
Subject words] Neoplasms;
Proteins;
Citrullination;
Two dimensional Western blot
- From:
Chinese Journal of Oncology
2015;(3):165-169
- CountryChina
- Language:Chinese
-
Abstract:
Objective The conversion of arginine into citrulline, termed citrullination, has important consequences for the structure and function of proteins.The present study aimed to identify novel citrullinated proteins in 10 tumor cell lines by 2-D Western blotting (2-D WB).Methods Two identical two-dimensional electrophoresis (2-DE) gels were prepared using extracts from ten cultured human tumor cell lines: ECA ( esophageal cancer cells ) , HEPG2 ( hepatocellular carcinoma cells ) , SKOV3 ( ovarian cancer cells),MCF-7 (breast cancer cells),H292 (lung mucoepidermoid carcinoma cells), HeLa (cervical cancer cells), Lovo (colon cancer cells), OS-RC (renal cell carcinoma cells), PANC-1 (pancreatic cancer cells), and SGC (gastric cancer cells).The expression profiles on one 2-DE gels were trans-blotted to PVDF membranes, and the blots were then probed with an anti-citrulline antibody.By comparing the 2-DE profile with the parallel 2-D WB profile at a global level, protein spots with immuno-signals were collected from the second 2-DE gel and identified using mass spectrometry.Immunoprecipitation was used to verify the expressionandcitrullinationofthetargetedproteinsinthetumorcelllines.Results 2-DWBandmass spectrometry identified citrullinated ENO1 (α-enolase), HSP60 (heat shock protein 60), KRT8 (keratin 8), TUBB (tubulin beta), TCRβ(T cell receptor βchain), VIME (vimentin) and PDI in these cell lines.Immunoprecipitation analyses verified the expression and citrullination of ENO1, HSP60, KRT8, and TUBB in the total protein lysates of the tumor cell lines.Conclusion The citrullination of proteins ENO1, HSP60, KRT8, and TUBB suggests a new mechanism in the tumorigenic process.
