Analysis of the catalytic site of the actin ADP-ribosylating Clostridium perfringens iota toxin
Analysis of the catalytic site of the actin ADP-ribosylating Clostridium perfringens iota toxin
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DOI：
10.1016/0014-5793(96)00052-x
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发表时间：
1996-02-19
期刊：
影响因子：
3.5
通讯作者：
Aktories, K
中科院分区：
文献类型：
--
作者：
vanDamme, J;Jung, M;Aktories, K
在[羰基-C-14]NAD 存在的情况下，通过紫外线照射对肌动蛋白 ADP-核糖基化产气荚膜梭菌 iota 毒素的酶成分进行亲和标记。通过CNBr裂解和随后用胰蛋白酶进行蛋白水解产生含有放射性标记的肽，其氨基酸序列为Gly-Ser-Pro-Gly-Ala-Tyr-Leu-Ser-Ala-Ile-Pro-Gly-Tyr-Ala-G-ly-X-Tyr-Glu-Val-Leu-Leu-Asn-His-Gly-Ser-Lys，对应于Gly-363区域通过产气荚膜梭菌 iota 毒素中的 Lys-388。质谱数据以及PTH-氨基酸分析结果与位于378位的谷氨酸侧链的修饰一致。因此，通过与其他ADP-核糖基转移酶类比可以得出结论，除了Glu-380之外，Glu-378可能在伯氏梭菌iota毒素的活性位点中发挥关键作用。
The enzyme component of the actin ADP-rihosylating Clostridium perfringens iota toxin was affinity labelled by UV irradiation in the presence of [carbonyl-C-14]NAD. A peptide containing the radiolabel was generated by CNBr cleavage and subsequent proteolysis with trypsin, Its amino acid sequence is Gly-Ser-Pro-Gly-Ala-Tyr-Leu-Ser-Ala-Ile-Pro-Gly-Tyr-Ala-G-ly-X-Tyr-Glu-Val-Leu-Leu-Asn-His-Gly-Ser-Lys corresponding with the region Gly-363 through Lys-388 in the C. perfringens iota toxin. Mass spectrometric data as well as the results of the PTH-amino acid analysis are in line with a modification of a glutamic acid side chain located at position 378. Therefore, in addition to Glu-380, as could be concluded by analogy with other ADP-ribosyltransferases, Glu-378 may play a pivotal role in the active site of C. peufringens iota toxin.