| Chapter title |
Production and Purification of Recombinant SUMOylated Proteins Using Engineered Bacteria.
|
|---|---|
| Chapter number | 4 |
| Book title |
SUMO
|
| Published in |
Methods in molecular biology, January 2016
|
| DOI | 10.1007/978-1-4939-6358-4_4 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-6356-0, 978-1-4939-6358-4
|
| Authors |
Frédérique Brockly, Marc Piechaczyk, Guillaume Bossis, Brockly, Frédérique, Piechaczyk, Marc, Bossis, Guillaume |
| Editors |
Manuel S. Rodriguez |
| Abstract |
SUMO is a ubiquitin-like protein that is covalently conjugated to numerous cellular proteins to modify their function and fate. Although large progresses have been made in the identification of SUMOylated proteins, the molecular consequences of their SUMOylation are generally unknown. This is, most often, due to the low abundance of SUMOylated proteins in the cell, usually less than 1 % of a given protein being modified at steady state. To gain insights into the role of specific SUMOylation targets, SUMO conjugation can be reconstituted in vitro using purified proteins. However, for most substrates, the efficiency of in vitro SUMOylation is too low to obtain sufficient amounts of their SUMOylated forms for biochemical studies. Here, we describe a detailed protocol to purify large amounts of recombinant SUMOylated proteins using bacteria modified to express His-tagged SUMO as well as the SUMO-activating and -conjugating enzymes. |
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 | 7 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Ph. D. Student | 2 | 29% |
| Lecturer | 1 | 14% |
| Student > Master | 1 | 14% |
| Researcher | 1 | 14% |
| Professor > Associate Professor | 1 | 14% |
| Other | 0 | 0% |
| Unknown | 1 | 14% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 3 | 43% |
| Agricultural and Biological Sciences | 2 | 29% |
| Immunology and Microbiology | 1 | 14% |
| Unknown | 1 | 14% |