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Related Experiment Video

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A highly selective mitochondria-targeting fluorescent K(+) sensor.

Xiangxing Kong1, Fengyu Su1, Liqiang Zhang1

  • 1Center for Biosignatures Discovery Automation, Biodesign Institute, Arizona State University, 1001 S. McAlister Ave., P.O. Box 876501, Tempe, AZ 85287 (USA).

Angewandte Chemie (International Ed. in English)
|August 25, 2015
PubMed
Summary

Researchers developed KS6, a novel fluorescent sensor for tracking mitochondrial potassium (K+) levels in live cells. This bright and selective sensor enables detailed studies of mitochondrial potassium dynamics during metabolic processes.

Keywords:
BODIPYfluorescent probesion channelsmitochondriapotassium

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Area of Science:

  • Biochemistry
  • Cell Biology
  • Biomedical Engineering

Background:

  • Intracellular potassium (K+) concentration is crucial for cellular metabolism.
  • Development of specific sensors for monitoring intracellular K+ dynamics, especially in mitochondria, remains limited.

Purpose of the Study:

  • To develop and characterize a highly selective fluorescent sensor (KS6) for monitoring mitochondrial potassium (K+) ion dynamics in live cells.

Main Methods:

  • Synthesis of KS6 by coupling triphenylphosphonium, borondipyrromethene (BODIPY), and triazacryptand (TAC).
  • Evaluation of KS6's response to K+ concentration, dynamic range, brightness, and selectivity (pH, metal ions).
  • Confocal microscopy for colocalization studies with MitoTracker Green in HeLa and U87MG cells.
  • Monitoring K+ flux in mitochondria upon stimulation with ionophores (nigericin, ionomycin).

Main Results:

  • KS6 demonstrated a wide response range (30–500 mM K+) with a large dynamic range (Fmax/F0 ≈130) and high brightness.
  • The sensor exhibited excellent selectivity, showing insensitivity to pH variations (5.5–9.0) and common metal ions.
  • Colocalization studies confirmed KS6's predominant localization within the mitochondria of live cells.
  • KS6 successfully detected mitochondrial K+ efflux and influx triggered by ionophores.

Conclusions:

  • KS6 is a highly selective, bright, and responsive fluorescent sensor for semiquantitative monitoring of mitochondrial potassium.
  • This sensor is suitable for investigating mitochondrial potassium flux and its role in cellular metabolic processes in live cells.