Screening of S-phase kinase-assuciated protein 2 in the cervical carcinoma HeLa cell
10.3872/j.issn.1007-385X.2018.03.008
- VernacularTitle:宫颈癌HeLa细胞内SKP2结合蛋白的筛选和功能预测
- Author:
JIA Jing
1
;
FANG Jianfei
1
;
REN Juan
1
;
JIA Zhenyu
1
;
WANG Xiaoju
1
Author Information
1. Center for Molecular Medicine,Academy of Medical Scinece of Zhejiang Province, Hangzhou 310013, Zhejiang, China
- Publication Type:Journal Article
- Keywords:
S-phase kinase-associated protein 2；co-immunoprecipitation；cervical cancer；binding-proteins
- From:
Chinese Journal of Cancer Biotherapy
2018;25(3):258-262
- CountryChina
- Language:Chinese
-
Abstract:
[Abstract] Objective： ：The co-immunoprecipitation and mass spectrometric analysis was carried out to obtain the S-phase kinase-associated protein 2 (SKP2)-binding proteins in HeLa cells, and the biological functions of these binding proteins were forecast. Methods: The co-immunoprecipitation system was established by co-immunoprecipitation and Western blotting assay; the specific protein gel of SKP2-binding proteins was obtained by SDS-PAGE and silver staining assay; the potential SKP2-binding proteins was identified by mass spectrometric analysis; and the GO (Gene ontology) analysis and KEGG analysis was carried out by bioinformatics technique. Results: The expression level of SKP2 protein in HeLa cells was high enough for co-immunoprecipitation assay; the co-immunoprecipitation system was established successfully, and SKP2-binding proteins was obtained; a total of 563 proteins were identified by mass spectrometric analysis, and 270 proteins with high credibility were obtained after screening. The GO analysis and KEGG analysis was carried out for the 270 proteins to forecast their functions and pathways. Conclusion: The SKP2-binding proteins were screened successfully, and it was the foundation for the subsequent screening of target-binding proteins and the search for targeting drugs.
- Full text:20180308.pdf
