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


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.

SUBMITTER: Iwasa C 

PROVIDER: S-EPMC3943108 | biostudies-literature | 2014 Jan

REPOSITORIES: biostudies-literature

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

Iwasa Chikako C   Tonozuka Takashi T   Shinoda Masaya M   Sagane Yoshimasa Y   Niwa Koichi K   Watanabe Toshihiro T   Yoshida Hiromi H   Kamitori Shigehiro S   Takao Toshifumi T   Oguma Keiji K   Nishikawa Atsushi A  

Acta crystallographica. Section F, Structural biology communications 20131224 Pt 1


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 col  ...[more]

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