Proteostasis: Methods and Protocols

ISBN-13: 9781493937547 / Angielski / Twarda / 2016 / 488 str.

Rune Matthiesen

Opis

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.

Spis treści

UPS Activation in the Battle against Aging and Aggregation-Related Diseases: An Extended Review.- Review And Literature Mining On Proteostasis Factors’ And Cancer.- Combining Zebrafish and Mouse Models to Test the Function of Deubiquitinating Enzyme (Dubs) Genes in Development:  Role Of Usp45 in the Retina.-  Immunodepletion and Immunopurification as Approaches for CSN Research.-  Studying Protein Ubiquitylation in Yeast.- Strategies to Detect Endogenous Ubiquitination of a Target Mammalian Protein.- In Vitro Ubiquitination – Self-Ubiquitination, Chain Formation, and Substrate Ubiquitination Assays.- Isolation of the Ubiquitin-Proteome From Tumour Cell Lines and Primary Cells Using Tubes.- Tubes-Mass Spectrometry for Identification and Analysis of the Ubiquitin-Proteome.- Isolation of Ubiquitinated Proteins to High Purity  from In Vivo Samples.-  Method for the Purification of Endogenous Unanchored Polyubiquitin Chains.- Fluorescent Tools for In Vivo Studies on the Ubiquitin-Proteasome System.- Bimolecular Fluorescence Complementation to Assay the Interactions of Ubiquitylation Enzymes in Living Yeast Cells.- Monitoring Ubiquitin-Coated Bacteria Via Confocal Microscopy.- Detection and Analysis of Cell Cycle-Associated Apc/C-Mediated Cellular Ubiquitylation In Vitro and In Vivo.-  Detection and Analysis Of Sumoylation Substrates In Vitro and In Vivo. - Detection of Protein-Protein Interactions and Post-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 and Cultured Cells.- Analysis Of Relevant Parameters For Autophagic Flux Using Hela Cells Expressing Egfp-Lc3.-  Analysis of Protein Oligomeric Species by Sucrose Gradients.- Analysis of Protein Oligomerization by Electrophoresis.-Blot-Ms of Carbonylated Proteins: A Tool to Identify Oxidized Proteins.- Quantitation of Protein Translation Rate In Vivo with Bioorthogonal Click-Chemistry.- A Simple Protocol for High Efficiency Protein Isolation after RNA Isolation from Mouse Thyroid and Other Very Small Tissue Samples.-  Monitoring Target Engagement Of Deubiquitylating Enzymes Using Activity Probes:  Past, Present, and Future.- Activity Based Profiling of Deubiquitylating Enzymes and Inhibitors in Animal Tissues.- High-Throughput Sirna Screening Applied to the Ubiquitin-Proteasome System.-  High-Throughput Yeast-Based Reporter Assay to Identify Compounds with Anti-Inflammatory Potential.- Using Alphascreen ^® to Identify Small Molecule Inhibitors Targeting a Conserved Host-Pathogen Interaction.- Global MS-Based Proteomics Drug Profiling.