Contents
Contents
Preface
Contributors
PART I BIOCHEMICAL METHODS TO STUDY THE UBIQUITIN PROTEASOME SYSTEM
1 Small-Molecule Inhibitors of Proteasome Activity
Maria Gaczynska and Pawel A. Osmulski
2 Purification of E1 and E1-like Enzymes
Art Haas
3 Assays for RING Family Ubiquitin Ligases
Manabu Furukawa, Paul Andrews, and Yue Xiong
4 Ubiquitin Chain Synthesis
Shahri Raasi and Cecile M. Pickart
5 Purification of Proteasomes, Proteasome Subcomplexes,
and Proteasome Associated Proteins from Budding Yeast
David S. Leggett, Michael H. Glickman, and Daniel Finley
6 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
7 Cell-Free Assay for Ubiquitin-Independent Proteasomal
Protein Degradation
Chaim Kahana and Yuval Reiss
8 Assays of Proteasome-Dependent Cleavage Products
Stefan Tenzer and Hansjörg Schild
9 Identification of Components of Protein Complexes
Carol E. Parker, Maria R. Warren, and Christoph H. Borchers
10 Mass Spectroscometric Determination of Protein Ubiqitination
Carol E. Parker, Viorel Mocanu, Susanna F. Greer,
and Christoph H. Borchers
11 Reconstitution of ER Associated Degradation
Using Yeast Membranes and Cytosol
Robert J. Lee, Ardythe A. McCracken, and Jeffrey L. Brodsky
12 Reticulocyte Lysate as a Model System to Study ER Membrane
Protein Degradation
Eric Carlson, Nathan Bays, Larry David, George Hilliard,
and William R. Skach
13 Deubiquitinating Enzyme Purifucation, Assay Inhibitors
and Characterization
Nathaniel S. Russell and Keith D. Wilkinson
PART II CELLULAR METHODS TO STUDY UBIQUITIN PROTEASOME DEPENDENT FUNCTIONS
14 Measuring Ubiquitin Conjugation in Cells
Edward G. Mimnaugh and Leonard M. Neckers
15 Assays for Proteasome Assembly and Maturation
R. Jürgen Dohmen, Markus K. London, Christoph Glanemann,
and Paula C. Ramos
16 N-terminal Ubiquitination
Aaron Ciechanover
17 Quantitating Defective Ribosome Products
Shu-Bing Qian, Jack R. Bennink, and Jonathan W. Yewdell
18 ER-Associated Quality Control and Degradation:
Screen for ERAD Mutants after EMS Mutagenesis
Antje Schaefer and Dieter H. Wolf
19 ER-Associated Quality Control and Degradation:
Genome Wide Screen for ERAD Components
Antje Schaefer and Dieter H. Wolf
20 Cystic Fibrosis Transmembrane Conducatance Regulator as a Model Substrate to Study ER Protein Quality
Control in Mammalian Cells
J. Michael Younger, Chun Yang Fan, Liling Chen,
Meredith F.N. Rosser, Andrea Fields, Hong-Yu Ren, Cam Patterson, and Douglas M. Cyr
21 Aggresome Formation
Michael J. Corboy, Philip J. Thomas, and W. Christian Wigley
22 Detection of Sumoylated Proteins
Kevin D. Sarge and Roland S. Hilgarth
23 Proteasome Inhibitors in Cancer Therapy
Robert Z. Orlowski
24 Parkinson Disease:
Assays for the Ubiquitin Ligase Activity of Neural Parkin
Michael G. Schlossmacher and Hideki Shimura
Index
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