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Full Description
This volume contains a collection of innovative techniques for studying targeted protein degradation. Chapters guide readers through heterobifunctional proteolysis-targeting chimeras (PROTACs) approaches, E3 ligase, E3 ligase-induced ubiquitylation, proteomic approaches, novel degrader molecules, molecular glue, and stabilize binding interaction between a target and E3 ubiquitin ligase. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and reagents, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols.
Authoritative and cutting-edge, Targeted Protein Degradation: Methods and Protocols aims to ensure successful results in this emerging field of drug discovery.
Contents
Protein Ligand Interactions Using Surface Plasmon Resonance.- High-Throughput Detection of Ligand-Protein Binding using a SplitLuc Cellular Thermal Shift Assay.- Proteins and their Interacting Partners: An Introduction to Protein-Ligand Binding Site Prediction Methods with a Focus on FunFOLD3.- Evaluating ligands for ubiquitin ligases using affinity beads.- Mechanistic and Structural Features of PROTAC Ternary Complexes.- MST and TRIC Technology to Reliably Study PROTAC Binary and Ternary Binding In Drug Development.- An in vitro Pull-down Assay of the E3 ligase:PROTAC:Substrate Ternary Complex to Identify Effective PROTACs.- Kinetic detection of E3: PROTAC: Target Ternary Complexes using NanoBRET technology in Live Cells.- Inducing Ubiquitylation and Protein Degradation as a Drug Development Strategy.- Methodological Versatility of Tandem Ubiquitin Binding Entities (TUBEs) for Understanding Poly-ubiquitination and its Various Chains.- Global Mass Spectrometry-Based Analysis of Protein Ubiquitination Using K-e-GG Remnant Antibody Enrichment.- Determination of Proteasomal Unfolding Ability.- Methods for Quantitative Assessment of Protein Degradation.- A High Throughput method to prioritize PROTAC intracellular target engagement and cell permeability using NanoBRET.- Profiling CELMoD Mediated Degradation of Cereblon Neosubstrates.- Global Proteome Profiling to Assess Changes in Protein Abundance using Isobaric Labeling and Liquid Chromatography Tandem Mass Spectrometry.- PHOTACs Enable Optical Control of Protein Degradation.- Protocols for synthesis of SNIPERs and the methods to evaluate the anti-cancer affects.
