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Screening of mammalian target of rapamycin inhibitors in natural product extracts by capillary electrophoresis in combination with high performance liquid chromatography-tandem mass spectrometry
Alternative Title毛细管电泳结合高效液相色谱-质谱联用用于天然产物中雷帕霉素靶标蛋白抑制剂的筛选
Zhang YM(张艳梅)1; Li F(李凤)1; LI MINGXIA1; Kang JW(康经武)1
2015
Source PublicationJ. Chromatogr. A
Volume1388Pages:267-273
AbstractIn this study, capillary electrophoresis (CE) combined with HPLC-MS/MS were used as a powerful platform for screening of inhibitors of mammalian target of rapamycin (mTOR) in natural product extracts. The screening system has been established by using 5-carboxyfluorescein labeled substrate peptide F-4EBP1, a known mTOR inhibitor AZD8055, and a small chemical library consisted of 18 natural product extracts. Biochemical screening of natural product extracts was performed by CE with laser induced fluorescence detection. The CE separation allowed a quantitative measurement of the phosphorylated product, hence the quantitation of enzymatic inhibition as well as inhibition kinetics. The hits are readily identified as long as the peak area of the phosphorylated product is reduced in comparison with the negative control. Subsequent assay-guided isolation of the active natural product extract was performed with HPLC-MS/MS to track the particular active components. The structures of the identified active components were elucidated by the molecular ions and fragmentaion information provided by MS/MS analysis. The CE-based assay method only requires minute pure compounds, which can be readily purified by HPLC. Therefore, the combination of CE and HPLC-MS/MS provides a high-throughput platform for screening bioactive compounds from the crude nature extracts. By taking the advantage of the screening system, salvianolic acid A and C in extract of Salvia miltiorrhiza were discovered as the new mTOR inhibitors. (C) 2015 Elsevier B.V. All rights reserved.
Subtype论文
Subject Area分析化学
DOI10.1016/j.chroma.2015.02.033
URL查看原文
Indexed BySCI
Language英语
WOS IDWOS:000351792400031
Citation statistics
Cited Times:14[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.sioc.ac.cn/handle/331003/39689
Collection分析化学研究室
Corresponding AuthorKang JW(康经武)
Affiliation1.中科院上海有机化学研究所, 生命有机化学国家重点实验室
2.上海科技大学
Recommended Citation
GB/T 7714
Zhang YM,Li F,LI MINGXIA,et al. Screening of mammalian target of rapamycin inhibitors in natural product extracts by capillary electrophoresis in combination with high performance liquid chromatography-tandem mass spectrometry[J]. J. Chromatogr. A,2015,1388:267-273.
APA 张艳梅,李凤,LI MINGXIA,&康经武.(2015).Screening of mammalian target of rapamycin inhibitors in natural product extracts by capillary electrophoresis in combination with high performance liquid chromatography-tandem mass spectrometry.J. Chromatogr. A,1388,267-273.
MLA 张艳梅,et al."Screening of mammalian target of rapamycin inhibitors in natural product extracts by capillary electrophoresis in combination with high performance liquid chromatography-tandem mass spectrometry".J. Chromatogr. A 1388(2015):267-273.
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