| Chapter title |
A Fluorescent In Vitro Assay to Investigate Paralog-Specific SUMO Conjugation.
|
|---|---|
| Chapter number | 5 |
| Book title |
SUMO
|
| Published in |
Methods in molecular biology, January 2016
|
| DOI | 10.1007/978-1-4939-6358-4_5 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-6356-0, 978-1-4939-6358-4
|
| Authors |
Nathalie Eisenhardt, Viduth K. Chaugule, Andrea Pichler, Eisenhardt, Nathalie, Chaugule, Viduth K., Pichler, Andrea |
| Editors |
Manuel S. Rodriguez |
| Abstract |
Protein modification with the small ubiquitin-related modifier SUMO is a potent regulatory mechanism implicated in a variety of biological pathways. In vitro sumoylation reactions have emerged as a versatile tool to identify and characterize novel SUMO enzymes as well as their substrates. Here, we present detailed protocols for the purification and fluorescent labeling of mammalian SUMO paralogs for their application in sumoylation assays. These assays provide a fast readout for in vitro SUMO chain formation activity of E3 ligases in a paralog-specific manner. Finally, we critically analyze the application of fluorescent SUMO proteins to study substrate modification in vitro revealing also the drawbacks of the system. |
X Demographics
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| United States | 1 | 100% |
Demographic breakdown
| Type | Count | As % |
|---|---|---|
| Members of the public | 1 | 100% |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 6 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Master | 2 | 33% |
| Researcher | 2 | 33% |
| Student > Ph. D. Student | 1 | 17% |
| Unknown | 1 | 17% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 3 | 50% |
| Veterinary Science and Veterinary Medicine | 1 | 17% |
| Agricultural and Biological Sciences | 1 | 17% |
| Unknown | 1 | 17% |