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This volume highlights the role of proteostasis in human health and associated disease model systems, reflecting its rising importance which has led to the development of new technologies to obtain insight into underling protein mechanistic events. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and cutting-edge, Proteostasis: Methods and Protocols aims to become a reference book on proteostasis in human health.
Sommario
UPS Activation in the Battle against Aging and Aggregation-RelatedDiseases: An Extended Review.- ReviewAnd Literature Mining On Proteostasis Factors' And Cancer.- Combining Zebrafish and Mouse Models to Test theFunction of Deubiquitinating Enzyme (Dubs) Genes in Development: Role Of Usp45 in the Retina.- Immunodepletion and Immunopurification as Approachesfor CSN Research.- Studying Protein Ubiquitylation in Yeast.- Strategies to Detect Endogenous Ubiquitination of aTarget Mammalian Protein.- In Vitro Ubiquitination - Self-Ubiquitination, ChainFormation, and Substrate Ubiquitination Assays.- Isolation of the Ubiquitin-Proteome From Tumour CellLines and Primary Cells Using Tubes.- Tubes-Mass Spectrometry for Identification andAnalysis of the Ubiquitin-Proteome.- Isolation of Ubiquitinated Proteins to HighPurity from In Vivo Samples.- Method for the Purification of Endogenous UnanchoredPolyubiquitin Chains.- Fluorescent Tools for In Vivo Studies onthe Ubiquitin-Proteasome System.- Bimolecular Fluorescence Complementation to Assaythe Interactions of Ubiquitylation Enzymes in Living Yeast Cells.- Monitoring Ubiquitin-Coated Bacteria Via ConfocalMicroscopy.- Detection and Analysis of Cell Cycle-AssociatedApc/C-Mediated Cellular Ubiquitylation In Vitro and In Vivo.- Detection and Analysis Of Sumoylation Substrates InVitro and In Vivo. - Detection of Protein-Protein Interactions andPost-Translational Modifications using the Proximity Ligation Assay:Application to the Study of the Sumo Pathway.- Dissecting of Sumo Dynamics by Mass Spectrometry.- Isolation of Lysosomes from Mammalian Tissues andCultured Cells.- Analysis Of Relevant Parameters For Autophagic FluxUsing Hela Cells Expressing Egfp-Lc3.- Analysis of Protein Oligomeric Species by SucroseGradients.- Analysis ofProtein Oligomerization by Electrophoresis.-Blot-Ms of Carbonylated Proteins: A Toolto Identify Oxidized Proteins.- Quantitation ofProtein Translation Rate In Vivo with Bioorthogonal Click-Chemistry.- A Simple Protocol for High Efficiency ProteinIsolation after RNA Isolation from Mouse Thyroid and Other Very Small TissueSamples.- Monitoring Target Engagement Of DeubiquitylatingEnzymes Using Activity Probes: Past,Present, and Future.- Activity Based Profiling of Deubiquitylating Enzymesand Inhibitors in Animal Tissues.- High-Throughput Sirna Screening Applied to theUbiquitin-Proteasome System.- High-Throughput Yeast-Based Reporter Assay toIdentify Compounds with Anti-Inflammatory Potential.- UsingAlphascreen ® to Identify Small Molecule Inhibitors Targeting aConserved Host-Pathogen Interaction.- Global MS-Based Proteomics Drug Profiling.
Riassunto
This volume highlights the role of proteostasis in human health and associated disease model systems, reflecting its rising importance which has led to the development of new technologies to obtain insight into underling protein mechanistic events.
