Chemical genetic control of protein levels: selective in vivo targeted degradation

J Am Chem Soc. 2004 Mar 31;126(12):3748-54. doi: 10.1021/ja039025z.

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Cell Line
  • Cysteine Endopeptidases / metabolism
  • Dihydrotestosterone / chemistry
  • Green Fluorescent Proteins
  • Humans
  • Ligands
  • Luminescent Proteins / genetics*
  • Luminescent Proteins / metabolism
  • Multienzyme Complexes / metabolism
  • Oligopeptides / chemical synthesis*
  • Oligopeptides / chemistry
  • Oligopeptides / pharmacology
  • Proteasome Endopeptidase Complex
  • Receptors, Androgen / genetics*
  • Receptors, Androgen / metabolism
  • Recombinant Fusion Proteins / genetics*
  • Recombinant Fusion Proteins / metabolism
  • Tacrolimus Binding Protein 1A / genetics*
  • Tacrolimus Binding Protein 1A / metabolism
  • Ubiquitins / metabolism

Substances

  • Ligands
  • Luminescent Proteins
  • Multienzyme Complexes
  • Oligopeptides
  • PROTAC-3
  • PROTAC-4
  • Receptors, Androgen
  • Recombinant Fusion Proteins
  • Ubiquitins
  • Dihydrotestosterone
  • Green Fluorescent Proteins
  • Cysteine Endopeptidases
  • Proteasome Endopeptidase Complex
  • Tacrolimus Binding Protein 1A