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Ubiquitin-Proteasome Protocols : Methods in Molecular Biology - Cam Patterson

Ubiquitin-Proteasome Protocols

Methods in Molecular Biology

By: Cam Patterson (Editor), Douglas M. Cyr (Editor)

Hardcover

Published: 1st April 2005
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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.

"...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

Prefacep. v
Contributorsp. ix
Biochemical Methods to Study the Ubiquitin-Proteasome System
Small-Molecule Inhibitors of Proteasome Activityp. 3
Purification of E1 and E1-Like Enzymesp. 23
Assays for RING Family Ubiquitin Ligasesp. 37
Ubiquitin Chain Synthesisp. 47
Purification of Proteasomes, Proteasome Subcomplexes, and Proteasome-Associated Proteins From Budding Yeastp. 57
Recognition and Processing of Misfolded Proteins by PA700, the 19S Regulatory Complex of the 26S Proteasomep. 71
Cell-Free Assay for Ubiquitin-Independent Proteasomal Protein Degradationp. 83
Assays of Proteasome-Dependent Cleavage Productsp. 97
Identification of Components of Protein Complexesp. 117
Mass Spectrometric Determination of Protein Ubiquitinationp. 153
Reconstitution of Endoplasmic Reticulum-Associated Degradation Using Yeast Membranes and Cytosolp. 175
Reticulocyte Lysate as a Model System to Study Endoplasmic Reticulum Membrane Protein Degradationp. 185
Deubiquitinating Enzyme Purification, Assay Inhibitors, and Characterizationp. 207
Cellular Methods to Study Ubiquitin-Proteasome-Dependent Functions
Measuring Ubiquitin Conjugation in Cellsp. 223
Assays for Proteasome Assembly and Maturationp. 243
N-Terminal Ubiquitinationp. 255
Quantitating Defective Ribosome Productsp. 271
Endoplasmic Reticulum-Associated Protein Quality Control and Degradation: Screen for ERAD Mutants After Ethylmethane Sulfonate Mutagenesisp. 283
Endoplasmic Reticulum-Associated Protein Quality Control and Degradation: Genome Wide Screen for ERAD Componentsp. 289
Cystic Fibrosis Transmembrane Conductance Regulator as a Model Substrate to Study Endoplasmic Reticulum Protein Quality Control in Mammalian Cellsp. 293
Aggresome Formationp. 305
Detection of Sumoylated Proteinsp. 329
Proteasome Inhibitors in Cancer Therapyp. 339
Parkinson's Disease: Assays for the Ubiquitin Ligase Activity of Neural Parkinp. 351
Indexp. 371
Table of Contents provided by Publisher. All Rights Reserved.

ISBN: 9781588292520
ISBN-10: 1588292525
Series: Methods in Molecular Biology
Audience: Professional
Format: Hardcover
Language: English
Number Of Pages: 381
Published: 1st April 2005
Publisher: Humana Press Inc.
Country of Publication: US
Dimensions (cm): 23.5 x 15.5  x 2.54
Weight (kg): 0.73