| Chapter title |
Simplified Enrichment of Plasma Membrane Proteins from Arabidopsis thaliana Seedlings Using Differential Centrifugation and Brij-58 Treatment
|
|---|---|
| Chapter number | 13 |
| Book title |
Brassinosteroids
|
| Published in |
Methods in molecular biology, January 2017
|
| DOI | 10.1007/978-1-4939-6813-8_13 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-6811-4, 978-1-4939-6813-8
|
| Authors |
Carina A. Collins, Michelle E. Leslie, Scott C. Peck, Antje Heese |
| Editors |
Eugenia Russinova, Ana I. Caño-Delgado |
| Abstract |
The plasma membrane (PM) forms a barrier between a plant cell and its environment. Proteins at this subcellular location play diverse and complex roles, including perception of extracellular signals to coordinate cellular changes. Analyses of PM proteins, however, are often limited by the relatively low abundance of these proteins in the total cellular protein pool. Techniques traditionally used for enrichment of PM proteins are time consuming, tedious, and require extensive optimization. Here, we provide a simple and reproducible enrichment procedure for PM proteins from Arabidopsis thaliana seedlings starting from total microsomal membranes isolated by differential centrifugation. To enrich for PM proteins, total microsomes are treated with the nonionic detergent Brij-58 to decrease the abundance of contaminating organellar proteins. This protocol combined with the genetic resources available in Arabidopsis provides a powerful tool that will enhance our understanding of proteins at the PM. |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 13 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Master | 3 | 23% |
| Student > Ph. D. Student | 2 | 15% |
| Other | 1 | 8% |
| Professor | 1 | 8% |
| Unspecified | 1 | 8% |
| Other | 3 | 23% |
| Unknown | 2 | 15% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 6 | 46% |
| Agricultural and Biological Sciences | 2 | 15% |
| Unspecified | 1 | 8% |
| Unknown | 4 | 31% |