Targeted chemical disruption of clathrin function in living cells

被引:27
作者
Moskowitz, HS
Heuser, J
McGraw, TE
Ryan, TA [1 ]
机构
[1] Cornell Univ, Weill Med Coll, Dept Biochem, New York, NY 10021 USA
[2] Washington Univ, Sch Med, Dept Cell Biol, St Louis, MO 63110 USA
关键词
D O I
10.1091/mbc.E03-04-0230
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
The accurate assignment of molecular roles in membrane traffic is frequently complicated by the lack of specific inhibitors that can work on rapid time scales. Such inhibition schemes would potentially avoid the complications arising from either compensatory gene expression or the complex downstream consequences of inhibition of an important protein over long periods (>12 h). Here, we developed a novel chemical tool to disrupt clathrin function in living cells. We engineered a cross-linkable form of clathrin by using an FK506-binding protein 12 (FKBP)-clathrin fusion protein that is specifically oligomerized upon addition of the cell-permeant cross-linker FK1012-A. This approach interrupts the normal assembly-disassembly cycle of clathrin lattices and results in a specific, rapid, and reversible similar to70% inhibition of clathrin function. This approach should be applicable to a number of proteins that must go through an assembly-disassembly cycle for normal function.
引用
收藏
页码:4437 / 4447
页数:11
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