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  Analyzing protein-phosphoinositide interactions with liposome flotation assays.

Busse, R. A., Scacioc, A., Schalk, A. M., Krick, R., Thumm, M., & Kühnel, K. (2016). Analyzing protein-phosphoinositide interactions with liposome flotation assays. In M. Waugh (Ed.), Lipid signaling protocols (2. ed., pp. 155-162). New York, N.Y.: Springer. doi:10.1007/978-1-4939-3170-5_13.

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Item Permalink: https://hdl.handle.net/11858/00-001M-0000-0028-4652-8 Version Permalink: https://hdl.handle.net/11858/00-001M-0000-002A-606D-1
Genre: Book Chapter

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2183338.pdf (Publisher version), 418KB
 
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http://link.springer.com/content/pdf/10.1007%2F978-1-4939-3170-5_13.pdf (Publisher version)
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 Creators:
Busse, R. A.1, Author           
Scacioc, A.1, Author           
Schalk, A. M.1, Author           
Krick, R., Author
Thumm, M., Author
Kühnel, K.2, Author           
Affiliations:
1Department of Neurobiology, MPI for biophysical chemistry, Max Planck Society, ou_578595              
2Research Group of Autophagy, MPI for Biophysical Chemistry, Max Planck Society, ou_1933285              

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Free keywords: Analytical ultracentrifuge; Hsv2; PROPPIN; Protein-lipid overlay assay; Small unilamellar vesicle s
 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.

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Language(s): eng - English
 Dates: 2015-112016
 Publication Status: Issued
 Pages: -
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 Table of Contents: -
 Rev. Type: Peer
 Identifiers: DOI: 10.1007/978-1-4939-3170-5_13
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Title: Lipid signaling protocols
Source Genre: Book
 Creator(s):
Waugh, M., Editor
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Publ. Info: New York, N.Y. : Springer, 2. ed.
Pages: XI, 257 Volume / Issue: - Sequence Number: - Start / End Page: 155 - 162 Identifier: ISBN: 978-1-4939-3169-9

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Title: Methods in Molecular Biology
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 Creator(s):
Walker, J. M., Editor
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Pages: - Volume / Issue: 1376 Sequence Number: - Start / End Page: - Identifier: -