Purification, crystallization and preliminary X-ray analysis of an HA17-HA70 (HA2-HA3) complex from Clostridium botulinum type C progenitor toxin

Acta Crystallogr F Struct Biol Commun. 2014 Jan;70(Pt 1):64-7. doi: 10.1107/S2053230X13032378. Epub 2013 Dec 24.

Abstract

The haemagglutinin (HA) complex of Clostridium botulinum type C toxin is composed of three types of subcomponents: HA33, HA17 and HA70 (also known as HA1, HA2 and HA3, respectively). Here, a 260 kDa HA17-HA70 complex was crystallized. His-tagged HA17 and maltose-binding-protein-tagged HA70 were expressed in Escherichia coli and their complex was affinity-purified using a combination of amylose resin chromatography and nickel-nitrilotriacetic acid agarose chromatography. Diffraction data were collected to 8.0 Å resolution and the crystal belonged to the tetragonal space group P4(1)2(1)2. The molecular-replacement solution indicated that one molecule of HA17 was bound to each HA70 monomer.

Keywords: Clostridium botulinum; haemagglutinin; lectins; neurotoxins.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Clostridium botulinum type C / chemistry*
  • Clostridium botulinum type C / isolation & purification*
  • Crystallization
  • Crystallography, X-Ray
  • Electrophoresis, Polyacrylamide Gel
  • Protein Subunits / chemistry*
  • Protein Subunits / isolation & purification*
  • Static Electricity

Substances

  • Protein Subunits