ADP-ribosylation factor 1 dependent clathrin-coat assembly on synthetic liposomes

Yunxiang Zhu, Matthew T. Drake, Stuart Kornfeld

Research output: Contribution to journalArticle

58 Scopus citations

Abstract

The assembly of clathrin-coated vesicles on Golgi membranes is initiated by the GTP-binding protein ADP ribosylation factor (ARF), which generates high-affinity membrane-binding sites for the heterotetrameric AP-1 adaptor complex. Once bound, the AP-1 recruits clathrin triskelia, which polymerize to form the coat. We have found that ARF·GTP also recruits AP-1 and clathrin onto protein-free liposomes. The efficiency of this process is modulated by the composition of the liposomes, with phosphatidylserine being the most stimulatory phospholipid. There is also a requirement for cytosolic factor(s) other than ARF. Thin-section electron microscopy shows the presence of clathrin-coated buds and vesicles that resemble those formed in vivo. These results indicate that AP-1-containing clathrin-coated vesicles can form in the absence of integral membrane proteins. Thus, ARF·GTP, appropriate lipids, and cytosolic factor(s) are the minimal components necessary for AP- 1 clathrin-coat assembly.

Original languageEnglish (US)
Pages (from-to)5013-5018
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume96
Issue number9
DOIs
StatePublished - Apr 27 1999

Keywords

  • AP-1 adaptor complex

ASJC Scopus subject areas

  • General

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