Chapter title |
Analyzing Protein–Phosphoinositide Interactions with Liposome Flotation Assays
|
---|---|
Chapter number | 13 |
Book title |
Lipid Signaling Protocols
|
Published in |
Methods in molecular biology, January 2016
|
DOI | 10.1007/978-1-4939-3170-5_13 |
Pubmed ID | |
Book ISBNs |
978-1-4939-3169-9, 978-1-4939-3170-5
|
Authors |
Ricarda A. Busse, Andreea Scacioc, Amanda M. Schalk, Roswitha Krick, Michael Thumm, Karin Kühnel |
Abstract |
Liposome flotation assays are a convenient tool to study protein-phosphoinositide interactions. Working with liposomes resembles physiological conditions more than protein-lipid overlay assays, which makes this method less prone to detect false positive interactions. However, liposome lipid composition must be well-considered in order to prevent nonspecific binding of the protein through electrostatic interactions with negatively charged lipids like phosphatidylserine. In this protocol we use the PROPPIN Hsv2 (homologous with swollen vacuole phenotype 2) as an example to demonstrate the influence of liposome lipid composition on binding and show how phosphoinositide binding specificities of a protein can be characterized with this method. |
Mendeley readers
Geographical breakdown
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Unknown | 24 | 96% |
Demographic breakdown
Readers by professional status | Count | As % |
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Researcher | 6 | 24% |
Student > Doctoral Student | 5 | 20% |
Student > Ph. D. Student | 5 | 20% |
Student > Master | 3 | 12% |
Student > Bachelor | 2 | 8% |
Other | 1 | 4% |
Unknown | 3 | 12% |
Readers by discipline | Count | As % |
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Unspecified | 1 | 4% |
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Computer Science | 1 | 4% |
Other | 2 | 8% |
Unknown | 3 | 12% |