期刊
JOURNAL OF CHROMATOGRAPHY A
卷 1562, 期 -, 页码 1-11出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.chroma.2018.05.034
关键词
Hsp 90 alpha; Mesoporous nanocomposites; Ligand Fishing; Real-time Imaging; Client proteins
资金
- National Natural Science Foundation of China [81573388]
- Qing Lan Project of Jiangsu Province
- Six talent peaks project of Jiangsu Province [YY-032]
- Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD)
Currently, nanosphere-based ligand fishing cannot be accomplished with imaging processing, although this step is important for real-time identification. Herein, a ligand fishing technique combined with real-time imaging is presented for the identification of ligands for heat shock protein 90 alpha (Hsp 90 alpha) from a complex matrix, Alisma plantago-aquatica Linn. crude extract, using Hsp 90 alpha-functionalized mesoporous silica nanoparticle (MSN)-InP/ZnS quantum dot (QD) nanocomposites as a support material. Twenty ligands for Hsp 90a were screened, and their structures were identified by mass spectrometry. The activities of the ligands were verified by real-time imaging of cells apoptotic morphological changes. Quantitative analysis showed that Alisma plantago-aquatica Linn contained 8.19 mu g/g Alisol F, which regarded as one typical component of Alisma plantago-aquatica Linn, and the extraction ratio of Alisol F was 76.2%. The precision for five replicate measurements was 7.0% (RSD). The prepared nanocomposites were also used to screen proteins from a mixture of cellular extracts, and five proteins from HeLa cells were identified as potential client proteins of Hsp 90 alpha. (C) 2018 Elsevier B.V. All rights reserved.
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