| Chapter title |
Yeast-Based Screening System for the Selection of Functional Light-Driven K+ Channels
|
|---|---|
| Chapter number | 17 |
| Book title |
Synthetic Protein Switches
|
| Published in |
Methods in molecular biology, March 2017
|
| DOI | 10.1007/978-1-4939-6940-1_17 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-6938-8, 978-1-4939-6940-1
|
| Authors |
Cristian Cosentino, Laura Alberio, Gerhard Thiel, Anna Moroni |
| Editors |
Viktor Stein |
| Abstract |
Ion channels control the electrical properties of cells by opening and closing (gating) in response to a wide palette of environmental and physiological stimuli. Endowing ion channels with the possibility to be gated by remotely applied stimuli, such as light, provides a tool for in vivo control of cellular functions in behaving animals. We have engineered a synthetic light-gated potassium (K(+)) channel by connecting an exogenous plant photoreceptor LOV2 domain to the K(+) channel pore Kcv. Here, we describe the experimental strategy that we have used to evolve the properties of the channel toward full control of light on pore gating. Our method combines rational and random mutagenesis of the channel followed by a yeast-based screening system for light-activated K(+) conductance. |
X Demographics
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Italy | 1 | 50% |
| United States | 1 | 50% |
Demographic breakdown
| Type | Count | As % |
|---|---|---|
| Members of the public | 2 | 100% |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 12 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Researcher | 5 | 42% |
| Student > Ph. D. Student | 2 | 17% |
| Student > Bachelor | 1 | 8% |
| Professor | 1 | 8% |
| Student > Master | 1 | 8% |
| Other | 1 | 8% |
| Unknown | 1 | 8% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 4 | 33% |
| Agricultural and Biological Sciences | 4 | 33% |
| Pharmacology, Toxicology and Pharmaceutical Science | 1 | 8% |
| Neuroscience | 1 | 8% |
| Unknown | 2 | 17% |