Stability of the Clostridium botulinum type A neurotoxin complex: an empirical phase diagram based approach

Mol Pharm. 2007 Jul-Aug;4(4):571-82. doi: 10.1021/mp0601244. Epub 2007 Jun 7.

Abstract

Clostridium botulinum type A neurotoxin (BoNT/A complex) is of great interest to the pharmaceutical industry. The drug itself is a natural complex of the toxin and a number of associated proteins. Surprisingly, relatively little is known about the exact structure and stability of the 900 kDa BoNT/A complex and its component proteins with the exception of the 150 kDa neurotoxin. In this study we describe the relative stability of the BoNT/A complex, the neurotoxin, and its associated proteins over a wide range of temperature and pH employing circular dichroism, intrinsic and 8-anilino-1-naphthalene sulfonate (ANS) fluorescence, and static light scattering. The data suggest a strong stabilizing effect of the associated proteins on the neurotoxin component. This data is compiled into empirical phase diagrams which permit the simultaneous visualization of multiple data sets over a wide range of conditions.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anilino Naphthalenesulfonates / metabolism
  • Circular Dichroism
  • Clostridium botulinum type A / chemistry*
  • Clostridium botulinum type A / isolation & purification
  • Clostridium botulinum type A / metabolism*
  • Drug Stability
  • Fluorescent Dyes / metabolism
  • Hydrogen-Ion Concentration
  • Light
  • Molecular Weight
  • Neurotoxins / chemistry*
  • Neurotoxins / isolation & purification
  • Neurotoxins / metabolism*
  • Scattering, Radiation
  • Spectrometry, Fluorescence
  • Temperature

Substances

  • Anilino Naphthalenesulfonates
  • Fluorescent Dyes
  • Neurotoxins
  • 1-anilino-8-naphthalenesulfonate