Steroid Receptor Methods

This volume of the Methods in Molecular Biology series is entirely devoted to the study of steroid receptor biology. Steroid hormone receptors represent a powerful system for the study of both the most fundamental molecular mec- nisms of gene regulation and control and the gross physiological responses of organisms to steroid hormones. Research in this field has brought forth advances in the treatment of cancer, endocrine disorders, and reproductive biology, and allowed elucidation of the fundamental biological mechanisms of gene expr- sion. In Steroid Receptor Methods: Protocols and Assays, the reader will find a collection of methods and protocols submitted by many fine steroid receptor researchers from throughout the world. These authors have been instructed to create a highly informative cross-section of the latest research techniques ava- able. The resulting work is timely, useful, and approachable for both the ex- rienced researcher and the novice to the field. Because the steroid receptor family is represented by a wonderfully diverse, yet strongly interrelated set of steroid receptor proteins, Steroid Receptor Methods contains protocols for the prod- tion and purification of a variety of receptor forms, including the progesterone, glucocorticoid, and androgen receptors. These procedures provide the raw ma- rial needed to conduct sophisticated biochemical analysis of receptor properties. Other techniques presented allow the reader to perform biochemical experiments on DNA binding characteristics, hormone binding assays, and protocols using combinatorial chemistry for drug discovery.

Includes supplementary material: sn.pub/extras

Klappentext
Research into steroid receptor biology holds great promise for the treatment of cancer and endocrine disorders and for elucidating the fundamental biological mechanisms of gene expression. In Steroid Receptor Methods: Protocols and Assays, a distinguished team of principal investigators and their associates describe in step-by-step detail a cross-section of the latest research techniques available for studying the endocrine system. As a basis for sophisticated biochemical analysis of receptor properties, the contributors provide methods for the production and purification of a variety of receptors, including progesterone, glucocorticoid, and androgen. Other protocols allow the reader to experiment with DNA-binding characteristics and hormone-binding assays and with the use of combinatorial chemistry for drug discovery. A series of novel methods utilizing the latest advances in immunochemistry, yeast two-hybrid screening, and fluorescence are included for the detection and analysis of a variety of cellular proteins that influence steroid receptor effectiveness. Finally, a discussion by example presents the most current bioinformatics tools and analytical techniques.
Timely and highly accessible, Steroid Receptor Methods: Protocols and Assays provides biochemists and molecular biologists studying endocrine systems with a full range of the powerful tools needed for elucidating protein interactions with receptors and DNA, as well as for drug discovery.

Inhalt
Bioinformatics.- Bioinformatics of Nuclear Receptors.- Phylogenetic Inference and Parsimony Analysis.- Purification Protocols.- Expression and Purification of Recombinant Human Progesterone Receptor in Baculovirus and Bacterial Systems.- High-Yield Purification of Functionally Active Glucocorticoid Receptor.- Production and Purification of Histidine-Tagged Dihydrotestosterone-Bound Full-Length Human Androgen Receptor.- Large Scale Production of Nuclear Receptor Ligand-Binding Domains.- Physical Structure of Nuclear Receptor-DNA Complexes.- Isolation of Steroid-Regulated Genes from the Uterus by mRNA Differential Display.- Identification of Nuclear Hormone Receptor Homologs by Screening Libraries with Highly Degenerate Oligonucleotide Probes.- Steroid Hormone-Binding Assays.- Use of [99mTc]Technetium-Labeled Steroids as Probes for Steroid Hormone Receptors.- Steroid Hormone Metabolites and Hormone Binding Assays.- Vitamin D3 Analog Screening.- Protein Interaction Assays.- Application of Green Fluorescent Protein to the Study of Dynamic Protein-Protein Interactions and Subcellular Trafficking of Steroid Receptors.- Knockout Mice and Steroid Receptor Research.- Yeast Two-Hybrid Screening for Proteins that Interact with Nuclear Hormone Receptors.- Isolation of a p300/CBP Cointegrator-Associated Protein Coactivator Complex.- Nonradioactive Photoaffinity Labeling of Steroid Receptors Using Western Blot Detection System.- Analysis of Steroid Hormone-Induced Histone Acetylation by Chromatin Immunoprecipitation Assay.- Analyzing the Contributions of Chromatin Structure in Nuclear Hormone Receptor Activated Transcription In Vivo.- Cotransfection Assays and Steroid Receptor Biology.- Estrogen Receptor mRNA In Situ Hybridization Using Microprobe System.- Cancer Research and Drug Discovery.- Solid Tumor Cancer Markers and Applications to Steroid Hormone Research.- Assessing Modulation of Estrogenic Activity of Environmental and Pharmaceutical Compounds Using MCF-7 Focus Assay.- Combinatorial Chemistry in Steroid Receptor Drug Discovery.- Identification of Nuclear Receptor Interacting Proteins Using Yeast Two- Hybrid Technology.