2. Academic Validation
  3. Mitochondria-targeted Probes for Imaging Protein Sulfenylation

Mitochondria-targeted Probes for Imaging Protein Sulfenylation

  • Sci Rep. 2018 Apr 27;8(1):6635. doi: 10.1038/s41598-018-24493-x.
Reetta J Holmila 1,  Stephen A Vance 2,  Xiaofei Chen 1,  Hanzhi Wu 1,  Kirtikar Shukla 1,  Manish S Bharadwaj 3,  Jade Mims 1,  Zack Wary 2,  Glen Marrs 4,  Ravi Singh 5,  Anthony J Molina 3,  Leslie B Poole 6,  S Bruce King 2,  Cristina M Furdui 7
Affiliations

Affiliations

  • 1 Department of Internal Medicine, Section on Molecular Medicine, Wake Forest University Health Sciences, Winston-Salem, NC, 27157, USA.
  • 2 Department of Chemistry, Wake Forest University, Winston-Salem, NC, 27109, USA.
  • 3 Department of Internal Medicine, Section on Gerontology and Geriatric Medicine, Wake Forest University Health Sciences, Winston-Salem, NC, 27157, USA.
  • 4 Department of Biology, Wake Forest University, Winston-Salem, NC, 27109, USA.
  • 5 Department of Cancer Biology, Wake Forest University Health Sciences, Winston-Salem, NC, 27157, USA.
  • 6 Department of Biochemistry, Wake Forest University Health Sciences, Winston-Salem, NC, 27157, USA.
  • 7 Department of Internal Medicine, Section on Molecular Medicine, Wake Forest University Health Sciences, Winston-Salem, NC, 27157, USA. [email protected].
PMID: 29703899 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.

Figures
Products