Ubiquitin-Proteasome Protocols

ISBN-13: 9781617374531 / Angielski / Miękka / 2010 / 381 str.

Cam Patterson; Douglas M. Cyr

Opis

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.

Recenzje

"...clearly written and thorough...a valuable addition to any lab entering, or already working in, the ubiquitin or proteasome field." - Doody's Health Science Book Review Journal

Spis treści

Part I. Biochemical Methods to Study the Ubiquitin-Proteasome System Small-Molecule Inhibitors of Proteasome Activity Maria Gaczynska and Pawel A. Osmulski Purification of E1 and E1-Like Enzymes Arthur L. Haas Assays for RING Family Ubiquitin Ligases Manabu Furukawa, Paul S. Andrews, and Yue Xiong Ubiquitin Chain Synthesis Shahri Raasi and Cecile M. Pickart Purification of Proteasomes, Proteasome Subcomplexes, and Proteasome-Associated Proteins From Budding Yeast David S. Leggett, Michael H. Glickman, and Daniel Finley Recognition and Processing of Misfolded Proteins by PA700, the 19S Regulatory Complex of the 26S Proteasome Chang-Wei Liu, Elizabeth Strickland, George N. DeMartino, and Philip J. Thomas Cell-Free Assay for Ubiquitin-Independent Proteasomal Protein Degradation Chaim Kahana and Yuval Reiss Assays of Proteasome-Dependent Cleavage Products Stefan Tenzer and Hansjörg Schild Identification of Components of Protein Complexes Carol E. Parker, Maria R. Warren, David R. Loiselle, Nedyalka N. Dicheva, and Christoph H. Borchers Mass Spectrometric Determination of Protein Ubiqitination Carol E. Parker, Viorel Mocanu, Maria R. Warren, Susanna F. Greer, Cameron O. Scarlett, and Christoph H. Borchers Reconstitution of Endoplasmic Reticulum-Associated Degradation Using Yeast Membranes and Cytosol Robert J. Lee, Ardythe A. McCracken, and Jeffrey L. Brodsky Reticulocyte Lysate as a Model System to Study Endoplasmic Reticulum Membrane Protein Degradation Eric Carlson, Nathan Bays, Larry David, and William R. Skach Deubiquitinating Enzyme Purification, Assay Inhibitors, and Characterization Nathaniel S. Russell and Keith D. Wilkinson Part II. Cellular Methods to Study Ubiquitin-Proteasome-Dependent Functions Measuring Ubiquitin Conjugation inCells Edward G. Mimnaugh and Leonard M. Neckers Assays for Proteasome Assembly and Maturation R. Jürgen Dohmen, Markus K. London, Christoph Glanemann, and Paula C. Ramos N-Terminal Ubiquitination, Aaron Ciechanover. Quantitating Defective Ribosome Products Shu-Bing Qian, Jack R. Bennink, and Jonathan W. Yewdell Endoplasmic Reticulum-Associated Protein Quality Control and Degradation: Screen for ERAD Mutants After Ethylmethane Sulfonate Mutagenesis Antje Schäfer and Dieter H. Wolf Endoplasmic Reticulum-Associated Protein Quality Control and Degradation: Genome Wide Screen for ERAD Components Antje Schäfer and Dieter H. Wolf Cystic Fibrosis Transmembrane Conductance Regulator as a Model Substrate to Study Endoplasmic Reticulum Protein Quality Control in Mammalian Cells J. Michael Younger, Chun-Yang Fan, Liling Chen, Meredith F. N. Rosser, Cam Patterson, and Douglas M. Cyr Aggresome Formation Michael J. Corboy, Philip J. Thomas, and W. Christian Wigley Detection of Sumoylated Proteins Roland S. Hilgarth and Kevin D. Sarge Proteasome Inhibitors in Cancer Therapy Robert Z. Orlowski Parkinson's Disease: Assays for the Ubiquitin Ligase Activity of Neural Parkin Michael G. Schlossmacher and Hideki Shimura Index

Blurb

The targeted addition of ubiquitin to proteins is now known to regulate many cellular events, with the enzymes involved in protein ubiquitination linked to a variety of clinical problems, such as cancer, heart disease, and immune dysfunction. In Ubiquitin-Proteasome Protocols, hands-on leaders in the field, many of whom originally developed the methods they describe, detail 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. The protocols follow the successful Methods in Molecular Biology™ series format, each offering step-by-step laboratory instructions, an introduction outlining the principle behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls.
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.