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Related Experiment Videos

Anthrax biosensor, protective antigen ion channel asymmetric blockade.

Kelly M Halverson1, Rekha G Panchal, Tam L Nguyen

  • 1United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Frederick, Maryland 21702-5011, USA.

The Journal of Biological Chemistry
|August 10, 2005
PubMed
Summary

Novel biosensors using anthrax protective antigen (PA63) nanopores can detect lethal toxin interactions and identify anthrax toxin complexes in plasma, aiding therapeutic and diagnostic development.

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Area of Science:

  • Biophysics
  • Microbiology
  • Toxicology

Background:

  • Biological agents like anthrax toxins pose significant threats, necessitating advanced methods for understanding toxin mechanisms and developing therapies.
  • Anthrax toxins comprise three proteins: protective antigen (PA), lethal factor (LF), and edema factor (EF).

Purpose of the Study:

  • To develop and validate a novel method for characterizing functional interactions of anthrax toxins using electrophysiological measurements.
  • To assess the potential of PA63 nanopore-based biosensors for anthrax diagnostics and therapeutics.

Main Methods:

  • Electrophysiological measurements of the heptameric PA63 ion channel.
  • Characterization of LF and EF interactions with the PA63 channel at pH 6.6.
  • Testing a neutralizing PA63 monoclonal antibody's effect on LF binding.

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  • Detection of anthrax lethal toxin complex in plasma samples from infected animals.

    • Main Results:

    • Full-length LF and EF altered the PA63 ion channel's current-voltage relationship, making it highly rectifying and diode-like.
    • The method confirmed that a neutralizing PA63 antibody blocks LF binding to the PA63 pore.
    • The biosensor technique successfully detected anthrax lethal toxin complex in infected animal plasma.

    Conclusions:

    • Electrophysiological analysis of PA63 nanopores offers a new approach to study functional toxin interactions.
    • PA63 nanopore-based biosensors show promise for real-time anthrax toxin detection, supporting therapeutic and diagnostic applications.