Lota toxin

Actin-ADP-ribosylating Toxins Cytotoxic Mechanisms of Clostridium botulinum C2 Toxin and Clostridium perfringens lota Toxin... 

Simpson LL, Stiles BG, Zapeda HH et al. (1987) Molecular basis for the pathological actions of Clostridium perfringens lota toxin. In Infect. Immun. 55 118-22 Simpson LL (1989) The binary toxin produced by Clostridium botulinum enters cells by receptor-mediated endocytosis to exert its pharmacologic effects. In J. Pharmacol. Exp. Ther. 251 1223-8... 

G-actin dissolved in G-buffer is ADP-ribosylated by addition of NAD (at least 10 iM, but NAD concentration should exceed actin concentration by about fivefold), 1 mM dithiothreitol, and 1. g/ml iota toxin (pH of NAD stock solution is to adjusted to 7.5 otherwise the pH of the reaction decreases). Incubation time is 30 to 60 min at 37°C (depending on actin concentration) or overnight on ice. Enzyme activity of iota toxin (i ) is only slighly decreased on ice. Thus, iota toxin-catalyzed ADP-ribosylation has to be stopped by removing the cosubstrate NAD or the toxin itself. NAD can be removed by gel filtration (e.g. by passing through a PD 10 column) or by dialysis against G-buffer. lota toxin is removed by immuno-precipitation with anti-iota toxin as descibed in 11.3.3. 

Probing the Actin Cytoskeleton by Clostridium botulinum C2 Toxin and Clostridium pedringens lota Toxin 137... 

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