Chapter title |
Native Gel Approaches in Studying Proteasome Assembly and Chaperones
|
---|---|
Chapter number | 16 |
Book title |
The Ubiquitin Proteasome System
|
Published in |
Methods in molecular biology, September 2018
|
DOI | 10.1007/978-1-4939-8706-1_16 |
Pubmed ID | |
Book ISBNs |
978-1-4939-8705-4, 978-1-4939-8706-1
|
Authors |
Jeroen Roelofs, Anjana Suppahia, Kenrick A. Waite, Soyeon Park, Roelofs, Jeroen, Suppahia, Anjana, Waite, Kenrick A., Park, Soyeon |
Abstract |
Proteasomes are complex molecular machines that consist of 66 subunits. The assembly of these complexes is highly coordinated in a process that requires at least ten proteasome-specific molecular chaperones. One of the challenges in studying assembly intermediates is their relatively low abundance as compared to the proteasome holoenzyme. Therefore, superior separating techniques are crucial for analyses of proteasomal complexes in general and studies in the assembly in particular. For this reason, native gel analyses have been abundantly used in studying proteasomes, as they provide a high resolution. Native gels are very versatile and can be used in various combinatorial approaches. In this chapter, we outline two approaches to prepare samples for native gels. The first is a yeast cryogrinding method and the second a core particle (CP)-base reconstitution approach. We describe the native gel electrophoresis, as well as various downstream analyses, including 2D native-SDS-PAGE. These techniques and approaches can all be used, often in parallel, to gain a variety of information about activity and composition of the complexes separated by native gel. The potential combined approaches are discussed in this review. |
X Demographics
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 1 | 100% |
Demographic breakdown
Type | Count | As % |
---|---|---|
Members of the public | 1 | 100% |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 22 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Student > Ph. D. Student | 8 | 36% |
Student > Doctoral Student | 2 | 9% |
Researcher | 2 | 9% |
Student > Bachelor | 1 | 5% |
Unknown | 9 | 41% |
Readers by discipline | Count | As % |
---|---|---|
Biochemistry, Genetics and Molecular Biology | 9 | 41% |
Medicine and Dentistry | 2 | 9% |
Immunology and Microbiology | 1 | 5% |
Agricultural and Biological Sciences | 1 | 5% |
Unknown | 9 | 41% |