title: Electrophoretic Mobility Shift Assay (EMSA) and Supershift Assay of Cytochrome P450 2B6 in Response to Estrogen.
abstract: Electrophoretic mobility shift assay (EMSA) is an invaluable tool to study interaction of proteins with DNA. Estrogens are major female hormones and modulate biological function through estrogen receptor (ER). ER regulates its target gene expression via the classical mechanism in which ER directly binds to its target gene promoter or the nonclassical mechanism involving tethering of ER to other transcription factors (such as AP-1 proteins). Here, we describe the EMSA to examine the nonclassical mechanism of ER action in regulation of a gene CYP2B6 by using competition and supershift assays.
authors.affiliation: Department of Internal Medicine, Rush University Medical Center, 1735 W. Harrison Street, Chicago, 60612, IL, USA.
authors.forename: Kwi Hye
authors.initials: KH
authors.lastname: Koh
authors.affiliation: Departments of Pharmacy Practice and Biopharmaceutical Sciences, College of Pharmacy, University of Illinois at Chicago, 833 S. Wood Street, Chicago, IL, 60612, USA. yjeong@uic.edu.
authors.forename: Hyunyoung
authors.initials: H
authors.lastname: Jeong
mesh_terms: D001665:Binding Sites; D001667:Binding, Competitive; D065702:Cytochrome P-450 CYP2B6 / Q000235:genetics / Q000378:metabolism*; D024202:Electrophoretic Mobility Shift Assay; D047628:Estrogen Receptor alpha / Q000819:agonists / Q000378:metabolism; D004967:Estrogens / Q000494:pharmacology*; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D056945:Hep G2 Cells; D022781:Hepatocytes / Q000187:drug effects* / Q000201:enzymology; D006801:Humans; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D018808:Transcription Factor AP-1 / Q000378:metabolism; D057188:Workflow
chemical_list: C506487:ESR1 protein, human; D047628:Estrogen Receptor alpha; D004967:Estrogens; D018808:Transcription Factor AP-1; C585599:CYP2B6 protein, human; D065702:Cytochrome P-450 CYP2B6
keywords: AP-1; CYP2B6; EMSA; Estrogen; Estrogen receptor; Supershift