Organelle-Specific Nitric Oxide Detection in Living Cells via HaloTag Protein Labeling

PLoS One. 2015 Apr 29;10(4):e0123986. doi: 10.1371/journal.pone.0123986. eCollection 2015.

Abstract

Nitric oxide (NO) is a membrane-permeable signaling molecule that is constantly produced, transferred, and consumed in vivo. NO participates and plays important roles in multiple biological processes. However, spatiotemporal imaging of NO in living cells is challenging. To fill the gap in currently used techniques, we exploited the versatility of HaloTag technology and synthesized a novel organelle-targetable fluorescent probe called HTDAF-2DA. We demonstrate the utility of the probe by monitoring subcellular NO dynamics. The developed strategy enables precise determination of local NO function.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alkylation
  • Bacteria / enzymology
  • Cell Survival
  • Chlorides / chemistry
  • Fluorescent Dyes / chemistry*
  • HeLa Cells
  • Hepatocytes
  • Humans
  • Hydrolases / chemistry
  • MCF-7 Cells
  • Microscopy, Fluorescence
  • Nitric Oxide / analysis*
  • Optical Imaging
  • Organelles / chemistry*
  • Staining and Labeling

Substances

  • Chlorides
  • Fluorescent Dyes
  • Nitric Oxide
  • Hydrolases
  • haloalkane dehalogenase

Grants and funding

This work is supported by the National Special Fund for State Key Laboratory of Bioreactor Engineering of China (2060204), 973 Program (2013CB531200), NSFC (31225008, 91313301, 31071260, 31170815, and 91013012), Specialized Research Fund for the Doctoral Program of Higher Education (20100074110010), Shanghai Science and Technology Commission (12JC1402900 and 11DZ2260600), Dawn Program of Shanghai Education Commission (11SG31), State Key Laboratory of Bioreactor Engineering, 111 Project (B07023), and Fundamental Research Funds for the Central Universities. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.