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学科主题: Biochemical Research Methods
题名: Identification of Novel 14-3-3 zeta Interacting Proteins by Quantitative Immunoprecipitation Combined with Knockdown (QUICK)
作者: Ge, Feng1; Li, Wen-Liang2; Bi, Li-Jun3; Tao, Sheng-Ce4; Zhang, Zhi-Ping5; Zhang, Xian-En5
通讯作者: Ge, F, Chinese Acad Sci, Inst Hydrobiol, Wuhan 430072, Hubei, Peoples R China
关键词: 14-3-3 zeta ; Quantitative immunoprecipitation combined with knockdown (QUICK) ; Multiple myeloma (MM) ; BCL2-associated athanogene 3 (BAG3) ; BCL2-associated X protein (BAX)
刊名: JOURNAL OF PROTEOME RESEARCH
发表日期: 2010-11-01
DOI: 10.1021/pr100616g
卷: 9, 期:11, 页:5848-5858
收录类别: SCI
文章类型: Article
部门归属: [Ge, Feng] Chinese Acad Sci, Inst Hydrobiol, Wuhan 430072, Hubei, Peoples R China; [Li, Wen-Liang] Tokai Univ, Sch Sci & Technol, Shibuya Ku, Tokyo 1510063, Japan; [Bi, Li-Jun] Chinese Acad Sci, Inst Biophys, Natl Lab Biomacromol, Beijing 100101, Peoples R China; [Tao, Sheng-Ce] Shanghai Jiao Tong Univ, Shanghai Ctr Syst Biomed, Shanghai 200240, Peoples R China; [Zhang, Zhi-Ping; Zhang, Xian-En] Chinese Acad Sci, Wuhan Inst Virol, State Key Lab Virol, Wuhan 430071, Peoples R China
WOS标题词: Science & Technology ; Life Sciences & Biomedicine
资助者: Chinese Academy of Sciences ; National Protein Research Fund [2009CB825100, 2006CB910902]; Ministry of Health of China [2009ZX10004-107, 2008ZX10003-005]; State Key Development Program for Basic Research of China [2010CB529205]; Japan Society for the Promotion of Science
类目[WOS]: Biochemical Research Methods
研究领域[WOS]: Biochemistry & Molecular Biology
摘要: The family of 14-3-3 proteins has emerged as critical regulators of diverse cellular responses under both physiological and pathological conditions. To gain insight into the molecular action of 14-3-3 zeta in multiple myeloma (MM), we performed a systematic proteomic analysis of 14-3-3 zeta-associated proteins. This analysis, recently developed by Matthias Mann, termed quantitative immunoprecipitation combined with knockdown (QUICK), integrates RNAi, SILAC, immunoprecipitation, and quantitative MS technologies. Quantitative mass spectrometry analysis allowed us to distinguish 14-3-3 zeta-interacting proteins from background proteins, resulting in the identification of 292 proteins in total with 95 novel interactions. Three 14-3-3 zeta-interacting proteins-BAX, HSP70, and BAG3-were further confirmed by reciprocal coimmunoprecipitations and colocalization analysis. Our results therefore not only uncover a large number of novel 14-3-3 zeta-associated proteins that possess a variety of cellular functions, but also provide new research directions for the study of the functions of 14-3-3 zeta. This study also demonstrated that QUICK is a useful approach to detect specific protein protein interactions with very high confidence and may have a wide range of applications in the investigation of protein complex interaction networks.
英文摘要: The family of 14-3-3 proteins has emerged as critical regulators of diverse cellular responses under both physiological and pathological conditions. To gain insight into the molecular action of 14-3-3 zeta in multiple myeloma (MM), we performed a systematic proteomic analysis of 14-3-3 zeta-associated proteins. This analysis, recently developed by Matthias Mann, termed quantitative immunoprecipitation combined with knockdown (QUICK), integrates RNAi, SILAC, immunoprecipitation, and quantitative MS technologies. Quantitative mass spectrometry analysis allowed us to distinguish 14-3-3 zeta-interacting proteins from background proteins, resulting in the identification of 292 proteins in total with 95 novel interactions. Three 14-3-3 zeta-interacting proteins-BAX, HSP70, and BAG3-were further confirmed by reciprocal coimmunoprecipitations and colocalization analysis. Our results therefore not only uncover a large number of novel 14-3-3 zeta-associated proteins that possess a variety of cellular functions, but also provide new research directions for the study of the functions of 14-3-3 zeta. This study also demonstrated that QUICK is a useful approach to detect specific protein protein interactions with very high confidence and may have a wide range of applications in the investigation of protein complex interaction networks.
关键词[WOS]: IN-VIVO ; CROSS-LINKING ; CANCER-CELLS ; MOLECULAR CHAPERONES ; SIGNALING PATHWAYS ; PROTEOMIC ANALYSIS ; OXIDATIVE STRESS ; DIMER FORMATION ; LIGAND-BINDING ; MYELOMA CELLS
语种: 英语
WOS记录号: WOS:000283810500033
ISSN号: 1535-3893
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.ihb.ac.cn/handle/342005/13741
Appears in Collections:水生生物分子与细胞生物学研究中心_期刊论文

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作者单位: 1.Chinese Acad Sci, Inst Hydrobiol, Wuhan 430072, Hubei, Peoples R China
2.Tokai Univ, Sch Sci & Technol, Shibuya Ku, Tokyo 1510063, Japan
3.Chinese Acad Sci, Inst Biophys, Natl Lab Biomacromol, Beijing 100101, Peoples R China
4.Shanghai Jiao Tong Univ, Shanghai Ctr Syst Biomed, Shanghai 200240, Peoples R China
5.Chinese Acad Sci, Wuhan Inst Virol, State Key Lab Virol, Wuhan 430071, Peoples R China

Recommended Citation:
Ge, Feng; Li, Wen-Liang; Bi, Li-Jun; Tao, Sheng-Ce; Zhang, Zhi-Ping; Zhang, Xian-En.Identification of Novel 14-3-3 zeta Interacting Proteins by Quantitative Immunoprecipitation Combined with Knockdown (QUICK),JOURNAL OF PROTEOME RESEARCH,2010,9(11):5848-5858
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