| Chapter title |
High-Throughput Fluorometric Assay for Membrane–Protein Interaction
|
|---|---|
| Chapter number | 14 |
| Book title |
Lipid Signaling Protocols
|
| Published in |
Methods in molecular biology, January 2016
|
| DOI | 10.1007/978-1-4939-3170-5_14 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-3169-9, 978-1-4939-3170-5
|
| Authors |
Wonhwa Cho, Hyunjin Kim, Yusi Hu |
| Abstract |
Membrane-protein interaction plays key roles in a wide variety of biological processes. To facilitate rapid and sensitive measurement of membrane binding of soluble proteins, we developed a fluorescence-based quantitative assay that is universally applicable to all proteins. This fluorescence-quenching assay employs fluorescence protein (FP)-tagged proteins whose fluorescence intensity is greatly decreased when they bind vesicles containing synthetic lipid dark quenchers, such as N-dimethylaminoazobenzenesulfonylphosphatidylethanolamine (dabsyl-PE). This simple assay can be performed with either a spectrofluorometer or a plate reader and optimized for different proteins with various combinations of FPs and quenching lipids. The assay allows rapid, sensitive, and accurate determination of lipid specificity and affinity for various lipid binding domains and proteins, and also high-throughput screening of small molecules that modulate membrane binding of proteins. |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 10 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Researcher | 3 | 30% |
| Professor | 2 | 20% |
| Other | 2 | 20% |
| Student > Master | 1 | 10% |
| Student > Ph. D. Student | 1 | 10% |
| Other | 0 | 0% |
| Unknown | 1 | 10% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 6 | 60% |
| Agricultural and Biological Sciences | 2 | 20% |
| Unknown | 2 | 20% |