The Nuclear Receptor Superfamily

1. Front Matter

   Pages i-x

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2. Introduction

   1. Nuclear Receptors: One Big Family

      • Iain J. McEwan

      Pages 3-18

3. Ligand Binding and Nuclear Receptor Turnover

   1. Methods for Measuring Ligand Dissociation and Nuclear Receptor Turnover in Whole Cells

      • Elizabeth M. Wilson

      Pages 21-33

   2. Flow Cytometry as a Tool for Measurement of Steroid Hormone Receptor Protein Expression in Leukocytes

      • Cherie L. Butts, Esther M. Sternberg

      Pages 35-50

   3. X-Ray Crystallography of Agonist/Antagonist-Bound Receptors

      • Ashley C.W. Pike

      Pages 51-66

4. Nuclear Localization and DNA Binding

   1. FRAP and FRET Methods to Study Nuclear Receptors in Living Cells

      • Martin E. van Royen, Christoffel Dinant, Pascal Farla, Jan Trapman, Adriaan B. Houtsmuller

      Pages 69-96

   2. Receptor-DNA Interactions: EMSA and Footprinting

      • Jason T. Read, Helen Cheng, Stephen C. Hendy, Colleen C. Nelson, Paul S. Rennie

      Pages 97-122

   3. Chromatin Immunoprecipitation (ChIP) Methodology and Readouts

      • Charles E. Massie, Ian G. Mills

      Pages 123-137

5. Nuclear Receptor—Co-Regulatory Protein Interactions

   1. Yeast-Based Reporter Assays for the Functional Characterization of Cochaperone Interactions with Steroid Hormone Receptors

      • Heather A. Balsiger, Marc B. Cox

      Pages 141-156

   2. High Throughput Analysis of Nuclear Receptor–Cofactor Interactions

      • Michael L. Goodson, Behnom Farboud, Martin L. Privalsky

      Pages 157-169

   3. Binding Affinity and Kinetic Analysis of Nuclear Receptor/Co-Regulator Interactions Using Surface Plasmon Resonance

      • Derek N. Lavery

      Pages 171-186

   4. Using RNA Interference to Study Protein Function

      • Carol D. Curtis, Ann M. Nardulli

      Pages 187-204

   5. Using Intrinsic Fluorescence Emission Spectroscopy to Study Steroid Receptor and Coactivator Protein Conformation Dynamics

      • Kate Watt, Iain J. McEwan

      Pages 205-218

6. Pathophysiological Analysis of Nuclear Receptor Function

   1. Development of Phosphorylation Site-Specific Antibodies to Nuclear Receptors

      • Inés Pineda Torra, Julia A. Staverosky, Susan Ha, Susan K. Logan, Michael J. Garabedian

      Pages 221-235

   2. Tissue-Selective Knockouts of Steroid Receptors: A Novel Paradigm in the Study of Steroid Action

      • Karel De Gendt, Guido Verhoeven

      Pages 237-261

   3. Methods for Identifying and Studying Genetic Alterations in Hormone-Dependent Cancers

      • Outi R. Saramäki, Kati K. Waltering, Tapio Visakorpi

      Pages 263-277

7. Back Matter

   Pages 279-281

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Acting principally to control patterns of gene expression, nuclear receptors play vital roles during embryonic development and in the regulation of metabolic and reproductive functions in adult life, which proves this superfamily of ligand-activated transcription factors to be a crucial part of biological life. In The Nuclear Receptor Superfamily: Methods and Protocols, expert researchers describe a range of molecular, structural and cell biological techniques currently used to investigate the structure-function of nuclear receptors, together with experimental approaches that may lead to new therapeutic strategies for treating nuclear receptor-associated diseases. Written in the highly successful Methods in Molecular Biology™ series format, the chapters in this volume contain brief introductions to the topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, as well as notes from the experts to highlight tips on troubleshooting and avoiding known pitfalls.

Cutting-edge and easy to use, The Nuclear Receptor Superfamily: Methods and Protocols provides beneficial and time-saving guidance for all those undertaking research in this ever-growing field of study.

• Chaperone
• Cloning
• DNA
• DNA binding
• Ligand binding
• Natural ligands
• Protein interactions
• Regulation
• Steroid hormones
• gene expression
• leukocytes
• molecular biology
• transcription

From the reviews:

"The primary focus is on cellular biology, biochemistry and molecular biology. … It is written for a wide range of basic science and clinical researchers interested in transcriptional control of gene expression … . Biomedical scientists and clinicians … will appreciate the wealth of new information presented by the authors, all experts in the field. … This is an outstanding contribution to the literature. … This carefully edited collection of protocols will be valued by all those interested in steroid hormones and nuclear receptors." (Bruce A. Fenderson, Doody’s Review Service, May, 2009)

“This book form part of the series Methods in Molecular Biology and describes methods commonly used to investigate the structural and functional relationship of nuclear receptors, as well as the physiopathology and potential pharmacologic agents for use in therapy of these disorders. … On consulting this book, researchers, students and professionals in the areas of biology and public health will have an opportunity to update their knowledge of experimental methods and applications of functional studies of nuclear receptors and their therapeutic implications.” (Rosario D. C. Hirata, Brazilian Journal of Pharmaceutical Science, Spring, 2010)

• School of Medical Sciences, University of Aberdeen, Aberdeen, UK

  Iain J. McEwan

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