title: Crystal structure of the C. perfringens alpha-toxin with the active site closed by a flexible loop region.
abstract: Clostridium perfringens biotype A strains are the causative agents of gas-gangrene in man and are also implicated as etiological agents in sudden death syndrome in young domestic livestock. The main virulence factor produced by these strains is a zinc-dependent, phosphatidylcholine-preferring phospholipase C (alpha-toxin). The crystal structure of alpha-toxin, at pH 7.5, with the active site open and therefore accessible to substrate has previously been reported, as has calcium-binding to the C-terminal domain of the enzyme at pH 4.7. Here we focus on conformation changes in the N-terminal domain of alpha-toxin in crystals grown at acidic pH. These changes result in both the obscuring of the toxin active site and the loss of one of three zinc ions from it. Additionally, this "closed" form contains a small alpha helix, not present in the open structure, which hydrogen bonds to both the N and C-terminal domains. In conjunction with the previously reported findings that alpha-toxin can exist in active and inactive forms and that Thr74Ile and Phe69Cys substitutions markedly reduced the haemolytic activity of the enzyme, our work suggests that these loop conformations play a critical role in the activity of the toxin.
authors.affiliation: Department of Crystallography, Birkbeck College, Malet Street, London WC1E 7HX, UK.
authors.forename: Julian T
authors.initials: JT
authors.lastname: Eaton
authors.forename: Claire E
authors.initials: CE
authors.lastname: Naylor
authors.forename: Angela M
authors.initials: AM
authors.lastname: Howells
authors.forename: David S
authors.initials: DS
authors.lastname: Moss
authors.forename: Richard W
authors.initials: RW
authors.lastname: Titball
authors.forename: Ajit K
authors.initials: AK
authors.lastname: Basak
mesh_terms: D001665:Binding Sites; D002118:Calcium / Q000378:metabolism; D003016:Clostridium perfringens / Q000201:enzymology*; D018360:Crystallography, X-Ray; D006860:Hydrogen Bonding; D006863:Hydrogen-Ion Concentration; D008958:Models, Molecular; D009068:Movement; D018583:Pliability; D017434:Protein Structure, Tertiary; D010738:Type C Phospholipases / Q000737:chemistry* / Q000378:metabolism; D015032:Zinc / Q000378:metabolism
chemical_list: D010738:Type C Phospholipases; D015032:Zinc; D002118:Calcium