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Recruitment of Coat Proteins to Liposomes and Peptidoliposomes

  • Sonja Huser
  • Gregor Suri
  • Pascal Crottet
  • Martin SpiessEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1270)

Abstract

Intracellular transport within the cell is generally mediated by membrane vesicles. Their formation is typically initiated by activation of small GTPases that then recruit cytosolic proteins to the membrane surface to form a coat, interact with cargo and accessory proteins, and deform the lipid bilayer to produce a transport vesicle. Liposomes proved to be a useful tool to study the molecular mechanisms of these processes in vitro. Here we describe the use of liposomes and peptidoliposomes presenting lipid-coupled cytosolic tails of cargo proteins for the in vitro analysis of the membrane recruitment of AP-1 adaptors in the process of forming AP-1/clathrin coats. AP-1 recruitment is mediated by the GTPase Arf1 and requires specific lipids and cargo signals. Interaction with cargo induces AP-1 oligomerization already in the absence of clathrin. Without cargo peptides, accessory proteins, such as amphiphysin 2, can be identified that stabilize AP-1 binding to liposomal membranes.

Key words

Amphiphysin Arf1 Clathrin adaptor protein Coat protein Liposome Membrane traffic Peptidoliposome Protein sorting 

Notes

Acknowledgments

Our work was supported by grant 31003A-144111 from the Swiss National Science Foundation.

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Sonja Huser
    • 1
  • Gregor Suri
    • 1
  • Pascal Crottet
    • 1
  • Martin Spiess
    • 1
    Email author
  1. 1.BiozentrumUniversity of BaselBaselSwitzerland

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