Just I, Hennessey E S, Drummond D R, Aktories K, Sparrow J C
Institut für Pharmakologie und Toxikologie, Universität des Saarlandes, Homburg, Germany.
Biochem J. 1993 Apr 15;291 ( Pt 2)(Pt 2):409-12. doi: 10.1042/bj2910409.
Purified Drosophila indirect-flight-muscle actin and arthrin, an actin-ubiquitin conjugate, were ADP-ribosylated by Clostridium botulinum C2 toxin and Clostridium perfringens iota toxin. Phalloidin treatment inhibited the ADP-ribosylation of Drosophila actin and arthrin. Like actin, the ADP-ribose-arthrin linkage was sensitive towards hydroxylamine treatment, indicating arginine as the amino acid acceptor. Actin translated in vitro from the indirect-flight-muscle-specific gene Act88F was ADP-ribosylated by C. botulinum C2 toxin and C. perfringens iota toxin. Actin from the R177Q mutant of Act88F translated in vivo was not ADP-ribosylated confirming Arg-177 as the ADP-ribose acceptor. Mutant L176M actin was modified by both toxins, indicating that amino acid 176 of actin does not define the substrate specificity of C. botulinum C2 toxin. Whereas the gene products of various C-terminal mutants of Act88F translated in vitro (E334K, V339I, E364K, G368E, R372H) were substrates for ADP-ribosylation by C. botulinum C2 toxin and by C. perfringens iota toxin, neither toxin modified the N-terminal O-12 deletion mutant.
纯化的果蝇间接飞行肌肌动蛋白和关节蛋白（一种肌动蛋白 - 泛素共轭物）被肉毒梭菌C2毒素和产气荚膜梭菌iota毒素进行了ADP核糖基化。鬼笔环肽处理抑制了果蝇肌动蛋白和关节蛋白的ADP核糖基化。与肌动蛋白一样，ADP - 核糖 - 关节蛋白连接对羟胺处理敏感，表明精氨酸是氨基酸受体。从间接飞行肌特异性基因Act88F体外翻译的肌动蛋白被肉毒梭菌C2毒素和产气荚膜梭菌iota毒素进行了ADP核糖基化。体内翻译的Act88F的R177Q突变体的肌动蛋白未被ADP核糖基化，证实Arg - 177是ADP - 核糖受体。突变体L176M被两种毒素修饰，表明肌动蛋白的第176位氨基酸并不决定肉毒梭菌C2毒素的底物特异性。而在体外翻译的Act88F的各种C末端突变体（E334K、V339I、E364K、G368E、R372H）的基因产物是肉毒梭菌C2毒素和产气荚膜梭菌iota毒素进行ADP核糖基化的底物，但两种毒素都未修饰N末端O - 12缺失突变体。
