| Chapter title |
Identification of Brassinosteroid Signaling Complexes by Coimmunoprecipitation and Mass Spectrometry
|
|---|---|
| Chapter number | 12 |
| Book title |
Brassinosteroids
|
| Published in |
Methods in molecular biology, January 2017
|
| DOI | 10.1007/978-1-4939-6813-8_12 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-6811-4, 978-1-4939-6813-8
|
| Authors |
Walter van Dongen, Luc van Heerde, Sjef Boeren, Sacco C. de Vries |
| Editors |
Eugenia Russinova, Ana I. Caño-Delgado |
| Abstract |
A combination of coimmunoprecipitation (coIP) of tagged proteins followed by protein identification and quantitation using Liquid Chromatography Mass Spectrometry/Mass Spectrometry (LCMS/MS) has proven to be a reliable method to qualitatively characterize membrane-bound receptor complexes from plants. Success depends on a range of parameters, such as abundance and stability of the complex and functionality of the tagged receptors, efficiency of the protein complex isolation procedure, MS equipment, and analysis software in use. In this Chapter, we focus on the use of one of the green fluorescent protein-tagged receptors of the SOMATIC EMBRYOGENESIS RECEPTOR-LIKE KINASE (SERK) family, of which SERK3, also known as BRASSINOSTEROID INSENSITIVE1 (BRI1) ASSOCIATED KINASE1 (BAK1), is a coreceptor of BRI1. Like BRI1 itself, SERK3 is a leucine-rich repeat receptor kinase (LRR RK) with a single-pass transmembrane domain. The latest updated laboratory protocol is presented as well as examples of data analysis and typical results obtained. Potential drawbacks of the procedure employed for plant membrane proteins will be pointed out. |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 5 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Unspecified | 1 | 20% |
| Student > Master | 1 | 20% |
| Student > Ph. D. Student | 1 | 20% |
| Professor > Associate Professor | 1 | 20% |
| Unknown | 1 | 20% |
| Readers by discipline | Count | As % |
|---|---|---|
| Agricultural and Biological Sciences | 2 | 40% |
| Biochemistry, Genetics and Molecular Biology | 1 | 20% |
| Unspecified | 1 | 20% |
| Unknown | 1 | 20% |