Description
Humana Press Ubiquitin-Proteasome Protocols by Cam Patterson, Douglas M. Cyr
A collection of cutting-edge techniques for studying ubiquitin-dependent protein degradation via the proteasome. The topics covered range broadly from basic biochemistry to cellular assays to discovery techniques using mass spectrometric analysis. These biochemical and cellular methods are necessary to explore the ubiquitin-proteasome system and ubiquitin-proteasome-dependent functions. State-of-the-art and user-friendly, Ubiquitin-Proteasome Protocols offers novice and experienced bench scientists alike a thorough compendium of readily reproducible techniques that will accelerate discovery, enhance productivity, and permit manipulation of the system for varied research purposes.show more Part I. Biochemical Methods to Study the Ubiquitin-Proteasome SystemSmall-Molecule Inhibitors of Proteasome ActivityMaria Gaczynska and Pawel A. OsmulskiPurification of E1 and E1-Like EnzymesArthur L. HaasAssays for RING Family Ubiquitin LigasesManabu Furukawa, Paul S. Andrews, and Yue XiongUbiquitin Chain SynthesisShahri Raasi and Cecile M. PickartPurification of Proteasomes, Proteasome Subcomplexes, and Proteasome-Associated Proteins From Budding YeastDavid S. Leggett, Michael H. Glickman, and Daniel FinleyRecognition and Processing of Misfolded Proteins by PA700, the 19S Regulatory Complex of the 26S ProteasomeChang-Wei Liu, Elizabeth Strickland, George N. DeMartino, and Philip J. ThomasCell-Free Assay for Ubiquitin-Independent Proteasomal Protein DegradationChaim Kahana and Yuval ReissAssays of Proteasome-Dependent Cleavage ProductsStefan Tenzer and Hansjoerg SchildIdentification of Components of Protein ComplexesCarol E. Parker, Maria R. Warren, David R. Loiselle, Nedyalka N. Dicheva, and Christoph H. BorchersMass Spectrometric Determination of Protein UbiqitinationCarol E. Parker, Viorel Mocanu, Maria R. Warren, Susanna F. Greer, Cameron O. Scarlett, and Christoph H. BorchersReconstitution of Endoplasmic Reticulum-Associated Degradation Using Yeast Membranes and CytosolRobert J. Lee, Ardythe A. McCracken, and Jeffrey L. BrodskyReticulocyte Lysate as a Model System to Study Endoplasmic Reticulum Membrane Protein DegradationEric Carlson, Nathan Bays, Larry David, and William R. SkachDeubiquitinating Enzyme Purification, Assay Inhibitors, and CharacterizationNathaniel S. Russell and Keith D. WilkinsonPart II. Cellular Methods to Study Ubiquitin-Proteasome-Dependent FunctionsMeasuring Ubiquitin Conjugation inCellsEdward G. Mimnaugh and Leonard M. NeckersAssays for Proteasome Assembly and MaturationR. Jurgen Dohmen, Markus K. London, Christoph Glanemann, and Paula C. RamosN-Terminal Ubiquitination, Aaron Ciechanover. Quantitating Defective Ribosome ProductsShu-Bing Qian, Jack R. Bennink, and Jonathan W. YewdellEndoplasmic Reticulum-Associated Protein Quality Control and Degradation: Screen for ERAD Mutants After Ethylmethane Sulfonate MutagenesisAntje Schafer and Dieter H. WolfEndoplasmic Reticulum-Associated Protein Quality Control and Degradation: Genome Wide Screen for ERAD ComponentsAntje Schafer and Dieter H. WolfCystic Fibrosis Transmembrane Conductance Regulator as a Model Substrate to Study Endoplasmic Reticulum Protein Quality Control in Mammalian CellsJ. Michael Younger, Chun-Yang Fan, Liling Chen, Meredith F. N. Rosser, Cam Patterson, and Douglas M. CyrAggresome FormationMichael J. Corboy, Philip J. Thomas, and W. Christian WigleyDetection of Sumoylated ProteinsRoland S. Hilgarth and Kevin D. SargeProteasome Inhibitors in Cancer TherapyRobert Z. OrlowskiParkinson's Disease: Assays for the Ubiquitin Ligase Activity of Neural ParkinMichael G. Schlossmacher and Hideki ShimuraIndex