UM  > 健康科學學院
Studying forkhead box protein A1-DNA interaction and ligand inhibition using gold nanoparticles, electrophoretic mobility shift assay, and fluorescence anisotropy
Aung K.M.M.4; New S.Y.4; Hong S.3; Sutarlie L.4; Lim M.G.L.3; Tan S.K.3; Cheung E.3; Su X.4
2014-03-01
Source PublicationAnalytical Biochemistry
ISSN00032697 10960309
Volume448Issue:1Pages:95-104
Abstract

Forkhead box protein 1 (FoxA1) is a member of the forkhead family of winged helix transcription factors that plays pivotal roles in the development and differentiation of multiple organs and in the regulation of estrogen-stimulated genes. Conventional analytical methods - electrophoretic mobility shift assay (EMSA) and fluorescence anisotropy (FA) - as well as a gold nanoparticles (AuNPs)-based assay were used to study DNA binding properties of FoxA1 and ligand interruption of FoxA1-DNA binding. In the AuNPs assay, the distinct ability of protein-DNA complex to protect AuNPs against salt-induced aggregation was exploited to screen sequence selectivity and determine the binding affinity constant based on AuNPs color change and absorbance spectrum shift. Both conventional EMSA and FA and the AuNPs assay suggested that FoxA1 binds to DNA in a core sequence-dependent manner and the flanking sequence also played a role to influence the affinity. The EMSA and AuNPs were found to be more sensitive than FA in differentiation of sequence-dependent affinity. With the addition of a spin filtration step, AuNPs assay has been extended for studying small molecular ligand inhibition of FoxA1-DNA interactions enabling drug screening. The results correlate very well with those obtained using FA. © 2013 Published by Elsevier Inc.

KeywordEmsa Fluorescent Anisotropic Forkhead Box Protein Gold Nanoparticles Protein-dna Interactions
DOI10.1016/j.ab.2013.11.017
URLView the original
Indexed BySCI
WOS Research AreaBiochemistry & Molecular Biology ; Chemistry
WOS SubjectBiochemical Research Methods ; Biochemistry & Molecular Biology ; Chemistry, Analytical
WOS IDWOS:000331678500015
Fulltext Access
Citation statistics
Cited Times [WOS]:3   [WOS Record]     [Related Records in WOS]
Document TypeJournal article
CollectionFaculty of Health Sciences
Affiliation1.Nanyang Technological University
2.Yong Loo Lin School of Medicine
3.A-Star, Genome Institute of Singapore
4.A-Star, Institute of Materials Research and Engineering
Recommended Citation
GB/T 7714
Aung K.M.M.,New S.Y.,Hong S.,et al. Studying forkhead box protein A1-DNA interaction and ligand inhibition using gold nanoparticles, electrophoretic mobility shift assay, and fluorescence anisotropy[J]. Analytical Biochemistry,2014,448(1):95-104.
APA Aung K.M.M..,New S.Y..,Hong S..,Sutarlie L..,Lim M.G.L..,...&Su X..(2014).Studying forkhead box protein A1-DNA interaction and ligand inhibition using gold nanoparticles, electrophoretic mobility shift assay, and fluorescence anisotropy.Analytical Biochemistry,448(1),95-104.
MLA Aung K.M.M.,et al."Studying forkhead box protein A1-DNA interaction and ligand inhibition using gold nanoparticles, electrophoretic mobility shift assay, and fluorescence anisotropy".Analytical Biochemistry 448.1(2014):95-104.
Related Services
Recommend this item
Bookmark
Usage statistics
Export to Endnote
Google Scholar
Similar articles in Google Scholar
[Aung K.M.M.]'s Articles
[New S.Y.]'s Articles
[Hong S.]'s Articles
Baidu academic
Similar articles in Baidu academic
[Aung K.M.M.]'s Articles
[New S.Y.]'s Articles
[Hong S.]'s Articles
Bing Scholar
Similar articles in Bing Scholar
[Aung K.M.M.]'s Articles
[New S.Y.]'s Articles
[Hong S.]'s Articles
Terms of Use
No data!
Social Bookmark/Share
All comments (0)
No comment.
 

Items in the repository are protected by copyright, with all rights reserved, unless otherwise indicated.