Liposome-binding assays to assess specificity and affinity of phospholipid-protein interactions

Magdalena M. Julkowska, Johanna M. Rankenberg, Christa Testerink

Research output: Chapter in Book/Report/Conference proceedingChapterAcademicpeer-review

22 Citations (Scopus)

Abstract

Protein-lipid interactions play an important role in cellular protein relocation, activation and signal transduction. The liposome-binding assay is a simple and inexpensive method to examine protein-lipid binding in vitro. The phospholipids used for liposome production are dried and hydrated. Subsequent extrusion of the phospholipid mixture ensures the production of large unilamellar vesicles (LUV) filled with raffinose. Those LUVs can be easily separated from the aqueous solution by centrifugation. By incubating a protein of interest with the LUVs and subsequent centrifugation steps, the bound protein fraction can be determined using Western Blot or Coomassie staining. This technique enables analysis of protein-lipid binding affinity and specificity.

Original languageEnglish
Title of host publicationPlant Lipid Signaling Protocols
EditorsTeun Munnik, Ingo Heilmann
Place of PublicationTotowa
PublisherSpringer – Humana Press
Pages261-271
Number of pages11
ISBN (Electronic)9781627034012
ISBN (Print)9781627034005
DOIs
Publication statusPublished - 21 Mar 2013
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume1009
ISSN (Print)1064-3745

Keywords

  • Binding affinity
  • Large unilamellar vesicle (LUV)
  • Liposome assay
  • Phosphatidic acid
  • Phospholipid binding
  • Phospholipid signaling

Fingerprint Dive into the research topics of 'Liposome-binding assays to assess specificity and affinity of phospholipid-protein interactions'. Together they form a unique fingerprint.

Cite this