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Main description:
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.
Contents:
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 Hansjoerg 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. Jurgen 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 Schafer and Dieter H. Wolf
Endoplasmic Reticulum-Associated Protein Quality Control and Degradation: Genome Wide Screen for ERAD Components
Antje Schafer 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
PRODUCT DETAILS
Publisher: Springer (Humana Press Inc.)
Publication date: November, 2010
Pages: 393
Weight: 581g
Availability: Available
Subcategories: Biochemistry
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