Solution Structures of Bacillus anthracis Protective Antigen Proteins Using Small Angle Neutron Scattering and Protective Antigen 63 Ion Channel Formation Kinetics

Toxins (Basel). 2021 Dec 11;13(12):888. doi: 10.3390/toxins13120888.

Abstract

We are studying the structures of bacterial toxins that form ion channels and enable macromolecule transport across membranes. For example, the crystal structure of the Staphylococcus aureus α-hemolysin (α-HL) channel in its functional state was confirmed using neutron reflectometry (NR) with the protein reconstituted in membranes tethered to a solid support. This method, which provides sub-nanometer structural information, could also test putative structures of the Bacillus anthracis protective antigen 63 (PA63) channel, locate where B. anthracis lethal factor and edema factor toxins (LF and EF, respectively) bind to it, and determine how certain small molecules can inhibit the interaction of LF and EF with the channel. We report here the solution structures of channel-forming PA63 and its precursor PA83 (which does not form channels) obtained with small angle neutron scattering. At near neutral pH, PA83 is a monomer and PA63 a heptamer. The latter is compared to two cryo-electron microscopy structures. We also show that although the α-HL and PA63 channels have similar structural features, unlike α-HL, PA63 channel formation in lipid bilayer membranes ceases within minutes of protein addition, which currently precludes the use of NR for elucidating the interactions between PA63, LF, EF, and potential therapeutic agents.

Keywords: A-B toxins; Bacillus anthracis; PA63; PA83; alpha-hemolysin; anthrax; bacterial toxin; endotoxin; ion channel; pore forming toxin; protective antigen; β-barrel.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigens, Bacterial / analysis*
  • Antigens, Bacterial / chemistry*
  • Bacillus anthracis / chemistry*
  • Bacterial Toxins / analysis*
  • Bacterial Toxins / chemistry*
  • Kinetics
  • Molecular Structure
  • Protective Agents / analysis*
  • Protective Agents / chemistry*
  • Scattering, Small Angle

Substances

  • Antigens, Bacterial
  • Bacterial Toxins
  • Protective Agents