Chemical Genetic Control of Protein Levels: Selective in Vivo Targeted Degradation

John S. Schneekloth, Fabiana N. Fonseca, Michael Koldobskiy, Amit Mandal, Raymond Deshaies, Kathleen Sakamoto, Craig M. Crews

Research output: Contribution to journalArticlepeer-review

Abstract

Genetic loss of function analysis is a powerful method for the study of protein function. However, some cell biological questions are difficult to address using traditional genetic strategies often due to the lack of appropriate genetic model systems. Here, we present a general strategy for the design and syntheses of molecules capable of inducing the degradation of selected proteins in vivo via the ubiquitin-proteasome pathway. Western blot and fluorometric analyses indicated the loss of two different targets: green fluorescent protein (GFP) fused with FK506 binding protein (FKBP12) and GFP fused with the androgen receptor (AR), after treatment with PROteolysis TArgeting Chimeric molecules (PROTACS) incorporating a FKBP12 ligand and dihydrotestosterone, respectively. These are the first in vivo examples of direct small molecule-induced recruitment of target proteins to the proteasome for degradation upon addition to cultured cells. Moreover, PROTAC-mediated protein degradation offers a general strategy to create "chemical knockouts," thus opening new possibilities for the control of protein function.

Original languageEnglish (US)
Pages (from-to)3748-3754
Number of pages7
JournalJournal of the American Chemical Society
Volume126
Issue number12
DOIs
StatePublished - Mar 31 2004
Externally publishedYes

ASJC Scopus subject areas

  • Catalysis
  • Chemistry(all)
  • Biochemistry
  • Colloid and Surface Chemistry

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