Department of Anesthesiology

You are here

Targeted chemical disruption of clathrin function in living cells.

TitleTargeted chemical disruption of clathrin function in living cells.
Publication TypeJournal Article
Year of Publication2003
AuthorsMoskowitz HS, Heuser J, McGraw TE, Ryan TA
JournalMol Biol Cell
Volume14
Issue11
Pagination4437-47
Date Published2003 Nov
ISSN1059-1524
KeywordsAnimals, Bicyclo Compounds, Heterocyclic, CHO Cells, Clathrin, Cloning, Molecular, Coated Pits, Cell-Membrane, Cricetinae, Cricetulus, Fluorescence Recovery After Photobleaching, Microscopy, Microscopy, Electron, Protein Binding, Tacrolimus, Tacrolimus Binding Proteins, Thiazoles, Thiazolidines
Abstract

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 approximately 70% 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.

DOI10.1091/mbc.E03-04-0230
Alternate JournalMol. Biol. Cell
PubMed ID14551251
PubMed Central IDPMC266763
Grant ListDK-57689 / DK / NIDDK NIH HHS / United States
GM-07739 / GM / NIGMS NIH HHS / United States
GM-29641 / GM / NIGMS NIH HHS / United States
GM-61925 / GM / NIGMS NIH HHS / United States
NS-24692 / NS / NINDS NIH HHS / United States