Application of a proteolysis/mass spectrometry method for investigating the effects of inhibitors on hydroxylase structure

J Med Chem. 2009 May 14;52(9):2799-805. doi: 10.1021/jm900285r.

Abstract

Limited proteolysis coupled to matrix-assisted laser desorption/ionization (MALDI) mass spectrometric analyses can be used to screen for compounds that alter protein structure by monitoring stabilizing/destabilizing effects with respect to the rate and nature of proteolysis. When applied to prolyl hydroxylase 2, a key enzyme involved in human oxygen sensing, the method efficiently revealed differential effects on proteolytic stability for structurally similar compounds and for different substrates.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Catalytic Domain / drug effects
  • Enzyme Inhibitors / metabolism
  • Enzyme Inhibitors / pharmacology*
  • Humans
  • Hypoxia-Inducible Factor 1, alpha Subunit / chemistry
  • Hypoxia-Inducible Factor 1, alpha Subunit / metabolism
  • Inhibitory Concentration 50
  • Mixed Function Oxygenases / antagonists & inhibitors
  • Mixed Function Oxygenases / chemistry*
  • Mixed Function Oxygenases / metabolism*
  • Models, Molecular
  • Peptide Fragments / metabolism
  • Protein Stability / drug effects
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Substances

  • Enzyme Inhibitors
  • Hypoxia-Inducible Factor 1, alpha Subunit
  • Peptide Fragments
  • Mixed Function Oxygenases