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

被引:61
作者
Zhu, YX [1 ]
Drake, MT [1 ]
Kornfeld, S [1 ]
机构
[1] Washington Univ, Sch Med, Dept Internal Med, St Louis, MO 63110 USA
关键词
AP-1 adaptor complex;
D O I
10.1073/pnas.96.9.5013
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
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.
引用
收藏
页码:5013 / 5018
页数:6
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