Crystallization and preliminary X‐ray analysis of a thiol‐activated cytolysin

[摘要]

We present the first reported crystallization of a member of the thiol-activated family of protein toxins. Perfringolysin O, a virulence factor of Clostridium perfringens, has been crystallized in two different forms by the hanging drop vapor diffusion method. In one form the toxin crystallizes with PEG 20000 in the orthorhombic space group C222₁ with cell dimensions of and the crystals diffract to beyond 2.5 Å resolution. In the second form the toxin crystallizes in a large variety of organic solvents including malt whisky. This crystal form belongs to the orthorhombic space group P222₁ with unit cell dimensions and with diffraction observed to 2.4 Å resolution.

[学科分类] 生物化学/生物物理

[关键词] Perfringolysin O;Thiol-activated cytolysin;Pore-forming toxin;Crystallization;X-ray diffraction;Bicine;N;N-bis[2-hydroxyethyl]-glycine;DTT;1;4-dithiothreitol;HEPES;4-(2-hydroxyethyl)-1-piperazineethanesulphonic acid;MES;2-[N-morpholino]-ethanesulfonic acid;MPD;2-methyl-3;4-pentanediol;PEG;polyethylene glycol;PFO;perfringolysin O;SDS-PAGE;sodium dodecyl sulfate-polyacrylamide gel electrophoresis;Tris-HCI;2-amino-2-hydroxymethylpropane-1;3-diol hydrochloride