Activity-Based Proteome Profiling Probes Based on Woodward's Reagent K with Distinct Target Selectivity

Angew Chem Int Ed Engl. 2016 Jun 27;55(27):7766-71. doi: 10.1002/anie.201602666. Epub 2016 May 9.

Abstract

Woodward's reagent K (WRK) is a reactive heterocyclic compound that has been employed in protein chemistry to covalently and unspecifically label proteins at nucleophilic amino acids, notably at histidine and cysteine. We have developed a panel of WRK-derived activity-based probes and show that surprisingly and unexpectedly, these probes are fairly selective for a few proteins in the human proteome. The WRK-derived probes show unique reactivity towards the catalytic N-terminal proline in the macrophage migration inhibitory factor (MIF) and can be used to label and, if equipped with a fluorophore, to image MIF activities in living cells.

Keywords: activity-based probes; fluorescent imaging; protein labeling; proteomics.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Catalysis
  • Chromatography, High Pressure Liquid
  • HeLa Cells
  • Humans
  • Isoxazoles / chemistry*
  • Macrophage Migration-Inhibitory Factors / antagonists & inhibitors
  • Macrophage Migration-Inhibitory Factors / chemistry
  • Macrophage Migration-Inhibitory Factors / metabolism*
  • Microscopy, Fluorescence
  • Proline / chemistry
  • Proteome / analysis
  • Proteome / chemistry*
  • RNA Interference
  • RNA, Small Interfering / metabolism
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Substances

  • Isoxazoles
  • Macrophage Migration-Inhibitory Factors
  • Proteome
  • RNA, Small Interfering
  • N-ethyl-5-phenylisoxazolium-3'-sulfonate
  • Proline