Small molecule-induced and cooperative enzyme assembly on a 14-3-3 scaffold

A. den Hamer, L.J.M. Lemmens, M.A.D. Nijenhuis, C. Ottmann, M. Merkx, T.F.A. de Greef, L. Brunsveld

Research output: Contribution to journalArticleAcademicpeer-review

14 Citations (Scopus)
387 Downloads (Pure)

Abstract

Scaffold proteins regulate cell signalling by promoting the proximity of putative interaction partners. Although they are frequently applied in cellular settings, fundamental understanding of them in terms of, amongst other factors, quantitative parameters has been lagging behind. Here we present a scaffold protein platform that is based on the native 14-3-3 dimeric protein and is controllable through the action of a small-molecule compound, thus permitting study in an in vitro setting and mathematical description. Robust small-molecule regulation of caspase-9 activity through induced dimerisation on the 14-3-3 scaffold was demonstrated. The individual parameters of this system were precisely determined and used to develop a mathematical model of the scaffolding concept. This model was used to elucidate the strong cooperativity of the enzyme activation mediated by the 14-3-3 scaffold. This work provides an entry point for the long-needed quantitative insights into scaffold protein functioning and paves the way for the optimal use of reengineered 14-3-3 proteins as chemically inducible scaffolds in synthetic systems.

Original languageEnglish
Pages (from-to)331–335
Number of pages5
JournalChemBioChem
Volume18
Issue number3
DOIs
Publication statusPublished - 1 Feb 2017

Keywords

  • 14-3-3 proteins
  • combinatorial inhibition
  • cooperative effects
  • dimerization
  • protein engineering
  • protein scaffolds
  • Caspase 3/metabolism
  • Mutagenesis, Site-Directed
  • Substrate Specificity
  • Small Molecule Libraries/chemistry
  • Caspase 9/chemistry
  • Recombinant Fusion Proteins/biosynthesis
  • Protein Engineering
  • Protein Structure, Quaternary
  • 14-3-3 Proteins/chemistry
  • Enzyme Activation
  • Dimerization

Fingerprint

Dive into the research topics of 'Small molecule-induced and cooperative enzyme assembly on a 14-3-3 scaffold'. Together they form a unique fingerprint.

Cite this