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题名: QUICK identification and SPR validation of signal transducers and activators of transcription 3 (Stat3) interacting proteins
作者: Zheng, Peng2; Zhong, Qiu3; Xiong, Qian2; Yang, Mingkun2; Zhang, Jia1, 2; Li, Chongyang2; Bi, Li-Jun1; Ge, Feng2
通讯作者: Bi, LJ (reprint author), Chinese Acad Sci, Natl Lab Biomacromol, Inst Biophys, Beijing 100101, Peoples R China
关键词: Quantitative immunoprecipitation ; combined with knockdown (QUICK) ; Stable isotope labeling with amino ; acids in cell culture (SILAC) ; Stat3 ; 14-3-3 zeta ; Multiple myeloma ; Surface plasmon resonance (SPR)
刊名: JOURNAL OF PROTEOMICS
发表日期: 2012-01-04
DOI: 10.1016/j.jprot.2011.10.020
卷: 75, 期:3, 页:1055-1066
收录类别: SCI
文章类型: Article
部门归属: [Zhang, Jia; Bi, Li-Jun] Chinese Acad Sci, Natl Lab Biomacromol, Inst Biophys, Beijing 100101, Peoples R China; [Zheng, Peng; Xiong, Qian; Yang, Mingkun; Zhang, Jia; Li, Chongyang; Ge, Feng] Chinese Acad Sci, Inst Hydrobiol, Wuhan 430072, Peoples R China; [Zhong, Qiu] AntiTB Res Inst Guangdong Prov, IBP ARI Joint Ctr Res TB, Guangzhou 510630, Guangdong, Peoples R China
WOS标题词: Science & Technology ; Life Sciences & Biomedicine
资助者: Chinese Academy of Sciences; National Basic Research Program of China (973 Program)[2012CB518700]; National Laboratory of Biomacromolecules
类目[WOS]: Biochemical Research Methods
研究领域[WOS]: Biochemistry & Molecular Biology
摘要: Signal transducers and activators of transcription 3 (Stat3) has been reported to be involved in the pathogenesis of various human diseases and is constitutively active in human multiple myeloma (MM) U266 cells. The Stat3-regulated mechanisms involved in these processes, however, are not fully defined. To further understand the regulation of Stat3 activity, we performed a systematic proteomic analysis of Stat3 interacting proteins in U266 cells. This analysis, termed quantitative immunoprecipitation combined with knockdown (QUICK), combines RNAi, stable isotope labeling with amino acids in cell culture (SILAC), immunoprecipitation, and quantitative MS. As a result, quantitative mass spectrometry analysis allowed us to distinguish specific Stat3 interacting proteins from background proteins and led to the identification of a total of 38 proteins. Three Stat3 interacting proteins - 14-3-3 zeta, PRKCB and Hsp90 - were further confirmed by reciprocal co-immunoprecipitations and surface plasmon resonance (SPR) analysis. Our results therefore not only uncover a number of Stat3 interacting proteins that possess a variety of cellular functions, but also provide new insight into the mechanisms that regulate Stat3 activity and function in MM cells. (C) 2011 Elsevier B.V. All rights reserved.
英文摘要: Signal transducers and activators of transcription 3 (Stat3) has been reported to be involved in the pathogenesis of various human diseases and is constitutively active in human multiple myeloma (MM) U266 cells. The Stat3-regulated mechanisms involved in these processes, however, are not fully defined. To further understand the regulation of Stat3 activity, we performed a systematic proteomic analysis of Stat3 interacting proteins in U266 cells. This analysis, termed quantitative immunoprecipitation combined with knockdown (QUICK), combines RNAi, stable isotope labeling with amino acids in cell culture (SILAC), immunoprecipitation, and quantitative MS. As a result, quantitative mass spectrometry analysis allowed us to distinguish specific Stat3 interacting proteins from background proteins and led to the identification of a total of 38 proteins. Three Stat3 interacting proteins - 14-3-3 zeta, PRKCB and Hsp90 - were further confirmed by reciprocal co-immunoprecipitations and surface plasmon resonance (SPR) analysis. Our results therefore not only uncover a number of Stat3 interacting proteins that possess a variety of cellular functions, but also provide new insight into the mechanisms that regulate Stat3 activity and function in MM cells. (C) 2011 Elsevier B.V. All rights reserved.
关键词[WOS]: MULTIPLE-MYELOMA CELLS ; CONSTITUTIVE ACTIVATION ; PHOSPHATASE 2A ; KINASE-C ; IN-VIVO ; PROSTATE-CANCER ; KNOCKDOWN QUICK ; GENE ONTOLOGY ; TARGETING PKC ; APOPTOSIS
语种: 英语
WOS记录号: WOS:000299719900027
ISSN号: 1874-3919
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.ihb.ac.cn/handle/342005/16763
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, Natl Lab Biomacromol, Inst Biophys, Beijing 100101, Peoples R China
2.Chinese Acad Sci, Inst Hydrobiol, Wuhan 430072, Peoples R China
3.AntiTB Res Inst Guangdong Prov, IBP ARI Joint Ctr Res TB, Guangzhou 510630, Guangdong, Peoples R China

Recommended Citation:
Zheng, Peng; Zhong, Qiu; Xiong, Qian; Yang, Mingkun; Zhang, Jia; Li, Chongyang; Bi, Li-Jun; Ge, Feng.QUICK identification and SPR validation of signal transducers and activators of transcription 3 (Stat3) interacting proteins,JOURNAL OF PROTEOMICS,2012,75(3):1055-1066
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