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Fast screening of ligand-protein interactions based on ligand-induced protein stabilization of gold nanoparticles
New S.Y.3; Aung K.M.M.3; Lim G.L.2; Hong S.2; Tan S.K.2; Lu Y.3; Cheung E.2; Su X.3
2014-03-04
Source PublicationAnalytical Chemistry
ISSN00032700 15206882
Volume86Issue:5Pages:2361-2370
Abstract

High throughput screening of small molecular weight (LMW) ligands for protein and sensitive determination of ligand-induced protein stabilization is an important task in drug discovery and in protein structural and functional genomics studies. In this study, gold nanoparticles (AuNPs) and their aggregation property are used to develop a rapid and less equipment intensive assay for screening the interactions between LMW ligands and transcription factors (TFs) and human serum albumin. The assay is based on the fact that the aggregation/discpersion status of AuNPs is very sensitive to the conformation of surrounding proteins, and when a LMW ligand binds to the proteins, it can enhance proteins' salt and thermal stability, and therefore the protective effect on AuNPs from aggregation. Two TFs, i.e. FoxA1 (Forkhead box A1) and AP-2γ (activating enhancer binding protein 2 gamma), and 14 compounds from an NCI compounds library and human serum albumin (HSA) and three known ligands (ibuprofen, warfarin, and phenytoin) are involved to demonstrate the concept and to prove its generality and robustness. With this AuNP method, two strong LMW binders are identified for FoxA1 and AP-2γ; ligand induced protein stabilization is determined. The results have been verified using surface plasmon resonance spectroscopy (SPR) and differential static light scattering (DSLS) techniques. Tryptophan fluorescent measurement is also conducted to provide further information on protein conformational change upon LMW ligand loading as can be observed from AuNPs' UV-vis spectra. FoxA1 and AP-2γ are pivotal in regulating the transcriptional activity of estrogen receptor alpha and controlling the expression of estrogen-responsive breast cancer cells. Identification of drug candidates targeting these two transcription factors could be an alternative in treating breast cancer, in particular those that have become endocrine resistance. © 2014 American Chemical Society.

DOI10.1021/ac404241y
URLView the original
Indexed BySCI
WOS Research AreaChemistry
WOS SubjectChemistry, Analytical
WOS IDWOS:000332494100016
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Cited Times [WOS]:10   [WOS Record]     [Related Records in WOS]
Document TypeJournal article
CollectionFaculty of Health Sciences
Affiliation1.University of Illinois at Urbana-Champaign
2.A-Star, Genome Institute of Singapore
3.A-Star, Institute of Materials Research and Engineering
Recommended Citation
GB/T 7714
New S.Y.,Aung K.M.M.,Lim G.L.,et al. Fast screening of ligand-protein interactions based on ligand-induced protein stabilization of gold nanoparticles[J]. Analytical Chemistry,2014,86(5):2361-2370.
APA New S.Y..,Aung K.M.M..,Lim G.L..,Hong S..,Tan S.K..,...&Su X..(2014).Fast screening of ligand-protein interactions based on ligand-induced protein stabilization of gold nanoparticles.Analytical Chemistry,86(5),2361-2370.
MLA New S.Y.,et al."Fast screening of ligand-protein interactions based on ligand-induced protein stabilization of gold nanoparticles".Analytical Chemistry 86.5(2014):2361-2370.
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