Small-molecule-mediated rescue of protein function by an inducible proteolytic shunt

Matthew R. Pratt, Edmund C. Schwartz, Tom W. Muir

Research output: Contribution to journalArticle

62 Scopus citations

Abstract

Controlling protein function through posttranslational manipulations has emerged as an attractive complementary technology to existing genetic systems. Often these methods involve developing pharmacological agents to probe protein function without the need to generate a unique compound for each protein family. One common strategy uses small molecules that act as chemical inducers of dimerization by mediating the interaction of two proteins. Herein we report the use of a chemical inducer of dimerization for the development of a posttranslational technology for the manipulation of protein function. This system, split ubiquitin for the rescue of function (SURF), places the complementation of genetically split ubiquitin under the control of rapamycin-induced dimerization of FK506-binding protein and FKBP12-rapamycin- binding protein. Before complementation a "degron" dooms a protein of interest for destruction by the proteasome. Addition of rapamycin results in a proteolytic shunt away from degradation by inducing ubiquitin complementation and cleavage of the protein of interest from the degron. Importantly, the native protein is rescued. We characterized this system with firefly luciferase and went on to apply it to members of three important classes of proteins: proteases (caspase-3), kinases (v-Src), and transcription factors (Smad3). This general strategy should allow for inducible rescue of a variety of proteins in such a way that their native structure and function are maintained.

Original languageEnglish (US)
Pages (from-to)11209-11214
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume104
Issue number27
DOIs
StatePublished - Jul 3 2007
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • General

Keywords

  • Degradation
  • Rescue of function

Fingerprint Dive into the research topics of 'Small-molecule-mediated rescue of protein function by an inducible proteolytic shunt'. Together they form a unique fingerprint.

Cite this