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学科主题: 基础医学
题名:
Antigen-subtracted 2-DE/MS strategy, a novel proteomic analysis platform
作者: Zhao, Peng2; Zhang, Lijuan2; Zhong, Weijian3; Ying, Xianping3; Huang, He4; Yao, Biyun2; Yuan, Zhun2; Fu, Juanling2; Wang, Yuedan1; Zhou, Zongcan2
关键词: Antigen subtraction ; Two-dimensional gel electrophoresis ; Q-TOF MS/MS ; anti-Benzo(a)pyrene-7,8-dihydrodiol-9,10-epoxide (anti-BPDE) ; anti-BPDE-transformed cells
刊名: ARCHIVES OF TOXICOLOGY
发表日期: 2011
DOI: 10.1007/s00204-010-0539-3
卷: 85, 期:1, 页:35-41
收录类别: SCI
文章类型: Article
WOS标题词: Science & Technology
类目[WOS]: Toxicology
研究领域[WOS]: Toxicology
关键词[WOS]: MANGANESE CHLORIDE ; CELLS
英文摘要:

In the present study, we developed a novel proteomic research strategy named antigen-subtracted 2-DE/MS strategy and applied it to comparative proteomic analysis of anti-benzo(a)pyrene-7,8-dihydrodiol-9,10epoxide-transformed human bronchial epithelial cell line (16HBE-C) and its parental cell line (16HBE) G0/G1 cells. Following pre-purification by ammonium sulfate precipitation, rabbit antibodies against 16HBE G0/G1 cells were coupled with protein A/G PLUS-agarose under the maximal coupling rate of about 50%. The agarose-antibody complex was then used in immunoprecipitation known as antigen subtraction. When the mass ratio of antibody to the sample was 2.5-3:1, the subtraction rates for 16HBE and 16HBE-C G0/G1 cell proteins were 44 and 34%, respectively. Both subtracted and unsubtracted samples were then subjected to the 2-DE resolution. In 16HBE-subtracted 2-DE maps, 315 protein spots were subtracted and 49 new protein spots were detected, whereas in 16HBE-C-subtracted 2-DE maps, 287 protein spots were subtracted and 33 new protein spots were detected. Taken together, antigen subtraction results in 65 new protein spots being allowed to be detected, therefore, makes it possible to get more information of the samples. Finally, 4 protein spots only detected in 16HBE-C-subtracted 2-DE maps were analyzed by the Q-TOF MS/MS, and successfully identified as U6 snRNA-associated Sm-like protein LSm3, 60S acidic ribosomal protein P1, Peroxiredoxin-6 and 60S acidic ribosomal protein P2. These proteins are involved in carcinogenesis, oxidation stress and protein synthesis. In conclusion, the antigen-subtracted 2-DE/MS strategy could reduce the complexities of protein samples and therefore, improve the resolution for the sample analysis.

语种: 英语
所属项目编号: 2002CB512906 ; 30471470 ; 30671781 ; 30771995
项目资助者: China National Key Basic Research and Development Program ; National Nature Science Foundation of China
WOS记录号: WOS:000286620900006
Citation statistics:
内容类型: 期刊论文
版本: 出版稿
URI标识: http://ir.bjmu.edu.cn/handle/400002259/56833
Appears in Collections:基础医学院_免疫学系_期刊论文

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作者单位: 1.Peking Univ, Hlth Sci Ctr, Dept Immunol, Beijing 100191, Peoples R China
2.Peking Univ, Hlth Sci Ctr, Dept Toxicol, Beijing 100191, Peoples R China
3.Shanghai Municipal Ctr Dis Control & Prevent, Dept Toxicol, Shanghai 200336, Peoples R China
4.Peking Univ, Hlth Sci Ctr, Dept Microbiol, Beijing 100191, Peoples R China

Recommended Citation:
Zhao, Peng,Zhang, Lijuan,Zhong, Weijian,et al. Antigen-subtracted 2-DE/MS strategy, a novel proteomic analysis platform[J]. ARCHIVES OF TOXICOLOGY,2011,85(1):35-41.
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