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Estrogen Receptors

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Cover of 'Estrogen Receptors'

Table of Contents

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    Book Overview
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    Chapter 1 The Estrogen Receptors: An Overview from Different Perspectives
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    Chapter 2 Estrogen Receptors
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    Chapter 3 The Use of Real-Time Reverse Transcription-PCR for Assessing Estrogen Receptor and Estrogen-Responsive Gene Expression.
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    Chapter 4 Bioinformatics Analysis of Estrogen-Responsive Genes.
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    Chapter 5 Electrophoretic Mobility Shift Assay (EMSA) and Supershift Assay of Cytochrome P450 2B6 in Response to Estrogen
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    Chapter 6 Chromatin Immunoprecipitation Assay to Identify Genomic Binding Sites of Regulatory Factors.
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    Chapter 7 Chromatin Immunoprecipitation with Estrogen Receptor 1 and the Promoter of Greb1 in TM4 Sertoli Cells.
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    Chapter 8 Chromatin Immunoprecipitation-Sequencing (ChIP-seq) for Mapping of Estrogen Receptor-Chromatin Interactions in Breast Cancer.
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    Chapter 9 RNA-Seq Experiment and Data Analysis.
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    Chapter 10 DNA Microarray Analysis of Estrogen-Responsive Genes.
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    Chapter 11 Shotgun Proteomics Analysis of Estrogen Effects in the Uterus Using Two-Dimensional Liquid Chromatography and Tandem Mass Spectrometry
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    Chapter 12 Assessment of Protein Expression by Proximity Ligation Assay in the Nonhuman Primate Endometrium, Placenta, and Fetal Adrenal in Response to Estrogen
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    Chapter 13 Colocalization of Estrogen Receptors with the Fluorescent Tamoxifen Derivative, FLTX1, Analyzed by Confocal Microscopy
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    Chapter 14 Live-Cell Imaging of the Estrogen Receptor by Total Internal Reflection Fluorescence Microscopy
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    Chapter 15 In Situ Hybridization of Estrogen Receptors α and β and GPER in the Human Testis
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    Chapter 16 Purification of Histone Lysine Methyltransferase SMYD2 and Co-Crystallization with a Target Peptide from Estrogen Receptor α.
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    Chapter 17 Gold Nanoparticle-Based Förster Resonance Energy Transfer (FRET) Analysis of Estrogen Receptor: DNA Interaction
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    Chapter 18 Analysis of Interaction of Estradiol with Estrogen Receptor by NMR Spectroscopy
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    Chapter 19 Application of Circular Dichroism Spectroscopy to the Analysis of the Interaction Between the Estrogen Receptor Alpha and Coactivators: The Case of Calmodulin
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    Chapter 20 Surface Plasmon Resonance Study of Cooperative Interactions of Estrogen Receptor α and Specificity Protein 1 with Composite DNA Elements
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    Chapter 21 Estrogen Receptors
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    Chapter 22 The Synonymous Ala87 Mutation of Estrogen Receptor Alpha Modifies Transcriptional Activation Through Both ERE and AP1 Sites
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    Chapter 23 Estrogen Receptors
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    Chapter 24 Use of Reporter Genes to Analyze Estrogen Response: The Transgenic Zebrafish Model
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    Chapter 25 Comparison of the Effects of the Selective Estrogen Receptor Modulators Ospemifene, Raloxifene, and Tamoxifen on Breast Tissue in Ex Vivo Culture
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    Chapter 26 Estrogen Receptor Agonists and Antagonists in the Yeast Estrogen Bioassay.
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    Chapter 27 Silencing Estrogen Receptor-α with siRNA in the Intact Rodent Brain
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    Chapter 28 Silencing Estrogen Receptor-β with siRNA in Cultured Cells
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    Chapter 29 Experimental Procedures for Demonstration of MicroRNA Mediated Enhancement of Functional Neuroprotective Effects of Estrogen Receptor Agonists.
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    Chapter 30 Expression Profiles of Estrogen-Regulated MicroRNAs in Breast Cancer Cells.
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    Chapter 31 Estradiol-Induced Transcriptional Regulation of Long Non-Coding RNA, HOTAIR.
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    Chapter 32 Detection and Functional Analysis of Estrogen Receptor α Phosphorylated at Serine 216 in Mouse Neutrophils
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    Chapter 33 Estrogen Receptors
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    Chapter 34 Detection of Endogenous Selective Estrogen Receptor Modulators such as 27-Hydroxycholesterol.
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    Chapter 35 Phytoestrogens Activate the Estrogen Receptor in HepG2 Cells
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    Chapter 36 Detection of the Phosphorylation of the Estrogen Receptor α as an Outcome of GPR30 Activation
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    Chapter 37 GPER Mediates Non-Genomic Effects of Estrogen.
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    Chapter 38 GPER/GPR30 Knockout Mice: Effects of GPER on Metabolism
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    Chapter 39 Analysis of G-Protein Coupled Receptor 30 (GPR30) on Endothelial Inflammation
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    Chapter 40 Atherosclerosis and Vascular Biologic Responses to Estrogens: Histologic, Immunohistochemical, Biochemical, and Molecular Methods
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    Chapter 41 Assessing Direct Vascular Actions of Estrogens
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    Chapter 42 Estrogen Receptors
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    Chapter 43 Estrogen Receptors
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    Chapter 44 Erratum to: Chapter 43 Regulation of Activation Induced Deaminase (AID) by Estrogen
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    Chapter 45 Erratum to: Chapter 4 Bioinformatics Analysis of Estrogen-Responsive Genes
Attention for Chapter 16: Purification of Histone Lysine Methyltransferase SMYD2 and Co-Crystallization with a Target Peptide from Estrogen Receptor α.
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Chapter title
Purification of Histone Lysine Methyltransferase SMYD2 and Co-Crystallization with a Target Peptide from Estrogen Receptor α.
Chapter number 16
Book title
Estrogen Receptors
Published in
Methods in molecular biology, January 2016
DOI 10.1007/978-1-4939-3127-9_16
Pubmed ID
Book ISBNs
978-1-4939-3126-2, 978-1-4939-3127-9
Authors

Jiang, Yuanyuan, Holcomb, Joshua, Spellmon, Nicholas, Yang, Zhe, Yuanyuan Jiang, Joshua Holcomb, Nicholas Spellmon, Zhe Yang

Abstract

Methylation of estrogen receptor α by the histone lysine methyltransferase SMYD2 regulates ERα chromatin recruitment and its target gene expression. This protocol describes SMYD2 purification and crystallization of SMYD2 in complex with an ERα peptide. Recombinant SMYD2 is overexpressed in Escherichia coli cells. After release from the cells by French Press, SMYD2 is purified to apparent homogeneity with multiple chromatography methods. Nickel affinity column purifies SMYD2 based on specific interaction of its 6×His tag with the bead-immobilized nickel ions. Desalting column is used for protein buffer exchange. Gel filtration column purifies SMYD2 based on molecular size. The entire purification process is monitored and analyzed by SDS-polyacrylamide gel electrophoresis. Crystallization of SMYD2 is performed with the hanging drop vapor diffusion method. Crystals of the SMYD2-ERα peptide complex are obtained by microseeding using seeding bead. This method can give rise to large size of crystals which are suitable for X-ray diffraction data collection. X-ray crystallographic study of the SMYD2-ERα complex can provide structural insight into posttranslational regulation of ERα signaling.

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Unknown 5 100%

Demographic breakdown

Readers by professional status Count As %
Student > Bachelor 2 40%
Researcher 1 20%
Unknown 2 40%
Readers by discipline Count As %
Biochemistry, Genetics and Molecular Biology 1 20%
Chemistry 1 20%
Neuroscience 1 20%
Unknown 2 40%
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