Mitochondria-targeted Probes for Imaging Protein Sulfenylation

Sci Rep. 2018 Apr 27;8(1):6635. doi: 10.1038/s41598-018-24493-x.

Abstract

Mitochondrial reactive oxygen species (ROS) are essential regulators of cellular signaling, metabolism and epigenetics underlying the pathophysiology of numerous diseases. Despite the critical function of redox regulation in mitochondria, currently there are limited methods available to monitor protein oxidation in this key subcellular organelle. Here, we describe compounds for imaging sulfenylated proteins in mitochondria: DCP-NEt2-Coumarin (DCP-NEt2C) and rhodamine-based DCP-Rho1. Side-by-side comparison studies are presented on the reactivity of DCP-NEt2C and DCP-Rho1 with a model protein sulfenic acid (AhpC-SOH) and mitochondrial localization to identify optimized experimental conditions for labeling and visualization of protein sulfenylation that would be independent of mitochondria membrane potential and would not impact mitochondrial function. These probes are applied to image mitochondrial protein sulfenylation under conditions of serum starvation and in a cell culture model of lung cancer exposed to ionizing radiation and silver nanoparticles, agents serving dual functions as environmental stressors and cancer therapeutics.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • A549 Cells
  • Cytological Techniques / methods*
  • Humans
  • Indicators and Reagents / chemical synthesis*
  • Mitochondria / metabolism*
  • Mitochondrial Proteins / metabolism*
  • Molecular Probes / chemical synthesis*
  • Oxidation-Reduction
  • Protein Processing, Post-Translational*
  • Staining and Labeling / methods*
  • Sulfenic Acids / metabolism

Substances

  • Indicators and Reagents
  • Mitochondrial Proteins
  • Molecular Probes
  • Sulfenic Acids