Targeted Protein Degradation
- 1st Edition, Volume 681 - February 6, 2023
- Imprint: Academic Press
- Editor: George Burslem
- Language: English
- Hardback ISBN:9 7 8 - 0 - 3 2 3 - 9 9 2 0 0 - 8
- eBook ISBN:9 7 8 - 0 - 3 2 3 - 9 9 2 0 1 - 5
Targeted Protein Degradation, Volume 680 in the Methods in Enzymology series, highlights new advances in the field with this new volume presenting interesting chapters on a… Read more
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Request a sales quoteTargeted Protein Degradation, Volume 680 in the Methods in Enzymology series, highlights new advances in the field with this new volume presenting interesting chapters on a variety of timely topics, with each. Each written by an international board of authors.
- 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 Targeted Protein Degradation
Biochemists, biophysicists, molecular biologists, analytical chemists, and physiologists
- Cover image
- Title page
- Table of Contents
- Copyright
- Contributors
- Preface
- Chapter One: High-efficiency knock-in of degradable tags (dTAG) at endogenous loci in cell lines
- Abstract
- 1: Introduction
- 2: Expected outcomes
- 3: Advantages
- 4: Limitations
- 5: Notes
- 6: Troubleshooting
- Acknowledgments
- Conflict of interest
- Author contributions
- References
- Chapter Two: High throughput E3 ligase degron binding assays for novel PROTAC ligand discovery
- Abstract
- 1: Introduction
- 2: Degrons for E3 ligase binding assays
- 3: Method: Evaluating degron peptide binding to E3 ligases
- 4: Application of protocol to other ligases
- 5: Conclusion
- Acknowledgments
- References
- Chapter Three: Synthesis of SNIPERs against BCR-ABL with kinase inhibitors and a method to evaluate their growth inhibitory activity derived from BCR-ABL degradation
- Abstract
- 1: Introduction
- 2: Design and synthesis of SNIPERs against BCR-ABL kinase
- 3: Biological assays of SNIPER(ABL)s
- 4: Summary and conclusion
- Acknowledgments
- COI statement
- References
- Chapter Four: Harnessing nanobodies for target protein degradation through the Affinity-directed PROtein Missile (AdPROM) system
- Abstract
- 1: Introduction
- 2: General method
- 3: Assessing antibiotic tolerance for individual cell lines
- 4: Generation of retroviruses for stable integration of AdPROM
- 5: Transduction of the AdPROM retroviruses onto target cells
- 6: Assessing mode of action for targeted protein degradation through AdPROM
- 7: Discussion, summary and conclusion
- Acknowledgments
- References
- Chapter Five: Monitoring PROTAC interactions in biochemical assays using Lumit immunoassays
- Abstract
- 1: Introduction
- 2: Monitoring target engagement between a small molecule and protein
- 3: Monitoring ternary complex formation
- 4: Summary and conclusions
- Acknowledgments
- References
- Chapter Six: The In-Cell Western immunofluorescence assay to monitor PROTAC mediated protein degradation
- Abstract
- 1: Introduction
- 2: A case studies using the ICW to evaluate CDK6 degraders
- 3: A case studies using the ICW to evaluate PARP1 degraders in SNU719 cells
- 4: Expected outcomes
- 5: Quantification and statistical analysis
- Acknowledgments
- References
- Chapter Seven: Cereblon target validation using a covalent inhibitor of neosubstrate recruitment
- Abstract
- 1: Introduction
- 2: Synthesis of the covalent cereblon inhibitor EM12-SO2F
- 3: Use of EM12-SO2F probe for target validation
- 4: Conclusions
- References
- Chapter Eight: Advancing targeted protein degrader discovery by measuring cereblon engagement in cells
- Abstract
- 1: Introduction
- 2: Generation of a stable cell line expressing NanoLuc®-CRBN
- 3: Synthesis of a BODIPY™-lenalidomide fluorescent tracer
- 4: Cellular NanoBRET™ CRBN engagement assay
- 5: Conclusions
- Acknowledgments
- Conflicts of interest
- References
- Chapter Nine: Quantitative measurement of PROTAC intracellular accumulation
- Abstract
- 1: Introduction
- 2: Measurement of BTK-PROTAC binding constant (Kd)
- 3: NanoBRET in cell target engagement assay
- 4: Deduction of relative intracellular accumulation coefficient (K'P,D)
- References
- Chapter Ten: The importance of controls in targeted protein degradation: Determining mechanism
- Abstract
- 1: Introduction
- 2: Hijacking the UPS: Proteolysis targeting chimeras
- 3: Alternative methods for TPD
- 4: PROTAC development and considerations
- 5: Importance of recruiting elements and ternary complex formation
- 6: Linker, length, and characterization
- 7: Tag-based approaches
- 8: Critical controls for targeted protein degradation
- References
- Chapter Eleven: Crystallization of VHL-based PROTAC-induced ternary complexes
- Abstract
- 1: Introduction
- 2: Before you begin
- 3: Key resources table
- 4: Materials and equipment
- 5: Step-by-step method details
- 6: Summary and conclusions
- 7: Optimization and troubleshooting
- Acknowledgment
- References
- Chapter Twelve: Targeted protein degradation through light-activated E3 ligase recruitment
- Abstract
- 1: Introduction
- 2: Materials and equipment
- 3: Step-by-step method details
- 4: Safety considerations and standards
- 5: Summary
- Acknowledgments
- References
- Chapter Thirteen: A heterobifunctional molecule system for targeted protein acetylation in cells
- Abstract
- 1: Introduction
- 2: General method and statistical analysis
- 3: Generation of AceTAG cell lines
- 4: AlphaScreen assay to monitor ternary complex formation and cellular target engagement assay using photoaffinity probes
- 5: Immunoprecipitation and immunoblot analysis of acetylated proteins
- 6: Quantitative mass spectrometry to monitor AceTAG-induced acetylation on a protein of interest
- 7: Proteome-wide acetylation assessment
- 8: Summary and conclusions
- References
- Edition: 1
- Volume: 681
- Published: February 6, 2023
- No. of pages (Hardback): 340
- No. of pages (eBook): 340
- Imprint: Academic Press
- Language: English
- Hardback ISBN: 9780323992008
- eBook ISBN: 9780323992015
GB
George Burslem
George M. Burslem is currently an Assistant Professor for Biochemistry and Biophysics at the Perelman School of Medicine
Department of Biochemistry and Biophysics and Department of Cancer Biology, University of Pennsylvania. He was previously a Postdoctoral Fellow at Yale University. He completed his PhD. in the University of Leeds, and his MSci in the University of Bristol
Prof. Burslem's lab is interested in developing chemical tools to understand and modulate lysine post-translational modifications, specifically acetylation and ubiquitination. His laboratory is particularly interested in novel pharmacological approaches to modulate post-translational modifications which regulate gene expression and protein stability, employing a multidisciplinary approach including synthetic chemistry, biochemistry, biophysics and cell biology to probe biological systems in cancer biology.
Affiliations and expertise
Assistant Professor of Biochemistry and Biophysics, Department of Biochemistry and Biophysics, Perelman School of Medicine, University of Pennsylvania, USARead Targeted Protein Degradation on ScienceDirect