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Steroid Biochemistry
- 1st Edition, Volume 689 - October 2, 2023
- Editor: Trevor M Penning
- Language: English
- Hardback ISBN:9 7 8 - 0 - 4 4 3 - 1 8 5 9 6 - 0
- eBook ISBN:9 7 8 - 0 - 4 4 3 - 1 8 5 9 7 - 7
Steroid Biochemistry, Volume 688 in the Methods of Enzymology series, highlights new advances in the field, containing chapters on a variety of timely topics, including Cyt… Read more
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Request a sales quoteSteroid Biochemistry, Volume 688 in the Methods of Enzymology series, highlights new advances in the field, containing chapters on a variety of timely topics, including Cytochrome P450 Enzyme, Steroidogenic P450s (CYP11A1, 17A1, 21A2, 11B1, and 11B5), Steroid 17 alpha-hydroxylase/17,20-lyase (Cytochrome P450 17A1), Enzymes of Estrogen Biosynthesis, Aromatase and Steroid Sulfatase, Estrogenic 17b-Hydroxysteroid Dehydrogenase, Hydroxysteroid Dehydrogenases (HSD), 3a-Hydroxyssteroid Dehydrogenase, Approaches to Measuring 3b-Hydroxysteroid Dehydrogenase Type 1, 3b-Hydroxysteroid Dehydrogenase Type 2, and much more.
- Provides the authority and expertise of leading contributors from an international board of authors
- Presents the latest release in Methods in Enzymology serials
- Updated release includes the latest information on Steroid Biochemistry
Biochemists, biophysicists, molecular biologists, analytical chemists, and physiologists
- Cover image
- Title page
- Table of Contents
- Series Page
- Copyright
- Contributors
- Preface
- Section 1 Cytochrome P450 enzymes
- Chapter One: Generation of human steroidogenic cytochrome P450 enzymes for structural and functional characterization
- Abstract
- Abbreviations
- 1 Introduction
- 2 General strategies
- 3 Equipment and reagents for the protocols
- 4 Protein engineering
- 5 Expression
- 6 Purification: Protein extraction from membranes
- 7 Purification: Immobilized metal affinity chromatography
- 8 Purification: Ion exchange chromatography
- 9 Purification: Size exclusion chromatography
- 10 Expected results
- 11 Storage and usage
- 12 Spectral analysis
- 13 Safety considerations and standards
- Acknowledgments
- References
- Chapter Two: Steroid 17α-hydroxylase/17, 20-lyase (cytochrome P450 17A1)
- Abstract
- 1 Introduction
- 2 Heterologous expression and purification of human P450 17A1
- 3 Assays
- 4 Assay methods
- 5 Issues in the P450 17A1 field
- Acknowledgments
- References
- Section 2 Enzymes of estrogen biosynthesis
- Chapter Three: Aromatase and steroid sulfatase from human placenta
- Abstract
- Abbreviations
- 1 Introduction
- 2 General method and statistical analysis
- 3 Purification of AROM from human placenta
- 4 Purification of STS from human placenta
- 5 Concluding remarks
- Acknowledgment
- References
- Section 3 Hydroxysteroid dehydrogenases (HSD)
- Chapter Four: Approaches to assessing 3β-hydroxysteroid dehydrogenase-1
- Abstract
- 1 Introduction
- 2 3βHSD1 Genotyping protocol
- 3 3βHSD1 intracellular and extracellular activity using mass spectrometry
- 4 Determining 3βHSD1 activity in cell lines using tritiated DHEA and HPLC analysis of conditioned medium
- References
- Chapter Five: In vitro methods to assess 11β-hydroxysteroid dehydrogenase type 1 activity
- Abstract
- 1 Introduction
- 2 Historical context of in vitro 11β-HSD1 enzyme activity assays
- 3 Project- and resource-based choice of 11β-HSD1 activity assay
- 4 11β-HSD1 enzyme activity assay using whole cell lysates
- 5 11β-HSD1 enzyme activity assay using microsomal preparations
- 6 Enzyme activity assay using purified 11β-HSD1
- 7 11β-HSD1 activity assay in intact cells
- 8 Alternative quantification methods
- 9 Safety considerations and standards of the described methods
- 10 Concluding remarks
- Acknowledgments
- References
- Chapter Six: In vitro methods to assess 11β-hydroxysteroid dehydrogenase type 2 activity
- Abstract
- 1 Introduction
- 2 Overview of in vitro 11β-HSD2 activity assays
- 3 Whole cell lysate enzyme activity assay using LC-MS/MS for quantification
- 4 11β-HSD2 activity assay using tissue homogenates
- 5 Alternative methods to assess 11β-HSD2 activity
- 6 Project and resource based choice of 11β-HSD2 activity assays
- 7 Biomarkers to assess 11β-HSD2 activity in preclinical and clinical studies
- 8 Safety considerations and standards of the described methods
- 9 Concluding remarks
- Acknowledgments
- References
- Chapter Seven: 17β-Hydroxysteroid dehydrogenases types 1 and 2: Enzymatic assays based on radiometric and mass-spectrometric detection
- Abstract
- 1 Introduction
- 2 General overview of methods
- 3 Equipment
- 4 Materials
- 5 Stock solutions
- 6 Overexpression of HSD17B1 and HSD17B2 in E. coli
- 7 Enzymatic assays
- 8 Expected outcomes
- 9 Quantification and statistical analysis
- 10 Other considerations
- Acknowledgements
- References
- Section 4 Steroid reductases
- Chapter Eight: Approach1es for evolutionary, biochemical, and structural analysis of bacterial steroid 5α-reductases
- Abstract
- 1 Introduction
- 2 General method and statistical analysis
- 3 Identification of PbSRD5A
- 4 Construct design and bacmid generation
- 5 Expression of PbSRD5A proteins
- 6 Purification of His-tagged PbSRD5A protein
- 7 Crystallization
- 8 Model building, data processing, and structure determination
- 9 Enzyme activity analysis of PbSRD5A
- Acknowledgments
- References
- Chapter Nine: Bacterial expression, purification, and reconstitution of human steroid 5α-reductases in phospholipid liposomes and nanodiscs
- Abstract
- 1 Introduction
- 2 Protein expression in Escherichia coli and purification
- 3 Reconstitution of the proteins in liposomes
- 4 Preparation of 5αR1 and 5αR2 nanodiscs
- 5 Steady-state kinetics
- Acknowledgement
- References
- Chapter Ten: Steroid 5β-reductase (AKR1D1): Purification and characterization
- Abstract
- 1 Introduction
- 2 General overview of methods
- 3 Equipment
- 4 Materials
- 5 Stock solutions
- 6 Protein purification procedure for His-tagged proteins
- 7 Other considerations
- 8 Characterization of recombinant protein
- References
- Chapter Eleven: Aldo keto-reductase family 1C members 1 through 4 recombinant enzyme purification and enzyme assay
- Abstract
- 1 Introduction
- 2 Equipment
- 3 Materials
- 4 Stock solutions
- 5 Procedure
- 6 Other considerations
- Acknowledgments
- References
- Section 5 Steroid conjugating enzymes
- Chapter Twelve: Human cytosolic steroid sulfotransferases: Versatile and rapid activity assays
- Abstract
- 1 Introduction
- 2 Characteristics of human steroid sulfotransferases
- 3 Material, equipment and reagents
- 4 Protocols
- 5 Safety considerations
- 6 Analysis and statistics
- 7 Optimization
- 8 Alternative methods
- 9 Pros and Cons of SULT radiolabeled assays
- 10 Summary
- Acknowledgments
- References
- Section 6 Steroid analytical chemistry
- Chapter Thirteen: High sensitivity LC-MS methods for quantitation of hydroxy- and keto-androgens
- Abstract
- 1 Introduction
- 2 Instruments and equipment
- 3 Materials and reagents
- 4 Stock solutions
- 5 Procedure
- 6 Other considerations
- Funding
- References
- Chapter Fourteen: Detection and quantification of UV-transparent keto-androgens by dinitrophenylhydrazine derivatization for discontinuous kinetic assays
- Abstract
- 1 Introduction
- 2 Materials and chemicals
- 3 Protocols
- 4 Other considerations
- Acknowledgments
- References
- Chapter Fifteen: Investigating the biosynthesis and metabolism of 11β-hydroxyandrostenedione
- Abstract
- 1 Introduction
- 2 Conversion assays in mammalian cell models
- 3 C19 and C11-oxy C19 UPC2–MS/MS: Method development and validation
- 4 Conclusion
- Acknowledgements
- References
- Chapter Sixteen: Ultrasensitive quantification of estrogens in serum and plasma by liquid chromatography-tandem mass spectrometry
- Abstract
- 1 Introduction
- 2 Materials
- 3 Before you begin—Ān overview of the general workflow for the analysis of estrogens in biological samples
- 4 Protocols
- 5 Conclusion
- Funding
- References
- No. of pages: 480
- Language: English
- Edition: 1
- Volume: 689
- Published: October 2, 2023
- Imprint: Academic Press
- Hardback ISBN: 9780443185960
- eBook ISBN: 9780443185977
TP
Trevor M Penning
Dr. Trevor Penning, the Thelma Brown and Henry Charles Molinoff Professor of Pharmacology, is a professor of Systems Pharmacology and Translational Therapeutics at the University of Pennsylvania and has been the Director of the Center of Excellence in Environmental Toxicology (CEET) since its founding in 2006. He formulated the strategic vision of the CEET from its inception and built the environmental health science identity of the CEET at the University, regional, and national level.
As Director of the CEET, Dr. Penning has focused the center’s research around the connections between human disease and environmental exposures and emphasized the importance of using this research to mitigate health impacts on vulnerable communities both locally and globally.
Throughout his time as Director, Dr. Penning has focused his research on the connections between human health and anthropogenic pollution. Air pollution is responsible for more than 200,000 deaths globally each year and has been identified as a known human carcinogen by the World Health Organization.
Affiliations and expertise
Department of Systems Pharmacology and Translational Therapeutics, University of Pennsylvania Perelman School of Medicine, USARead Steroid Biochemistry on ScienceDirect