Targeted Chemical Disruption of Clathrin Function in Living Cells

Howard S. Moskowitz, John Heuser, Timothy E. McGraw, Timothy A. Ryan

Research output: Contribution to journalArticle

50 Citations (Scopus)

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 ∼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.

Original languageEnglish (US)
Pages (from-to)4437-4447
Number of pages11
JournalMolecular Biology of the Cell
Volume14
Issue number11
DOIs
StatePublished - Nov 2003
Externally publishedYes

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Clathrin
Tacrolimus Binding Proteins
Proteins
Gene Expression
Membranes

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

Cite this

Targeted Chemical Disruption of Clathrin Function in Living Cells. / Moskowitz, Howard S.; Heuser, John; McGraw, Timothy E.; Ryan, Timothy A.

In: Molecular Biology of the Cell, Vol. 14, No. 11, 11.2003, p. 4437-4447.

Research output: Contribution to journalArticle

Moskowitz, Howard S. ; Heuser, John ; McGraw, Timothy E. ; Ryan, Timothy A. / Targeted Chemical Disruption of Clathrin Function in Living Cells. In: Molecular Biology of the Cell. 2003 ; Vol. 14, No. 11. pp. 4437-4447.
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