产气荚膜梭菌iota毒素肌动蛋白ADP核糖基化催化位点的分析。
Analysis of the catalytic site of the actin ADP-ribosylating Clostridium perfringens iota toxin.
作者信息
van Damme J, Jung M, Hofmann F, Just I, Vandekerckhove J, Aktories K
机构信息
Institute for Biotechnology, Department of Biochemistry, Faculty of Medicine, University of Gent, Belgium.
出版信息
FEBS Lett. 1996 Feb 19;380(3):291-5. doi: 10.1016/0014-5793(96)00052-x.
The enzyme component of actin ADP-ribosylating Clostridium perfringens iota toxin was affinity labelled by UV irradiation in the presence of [carbonyl-14C]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-Gly-X-Tyr-Glu-Va l-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 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. perfringens iota toxin.
在[羰基 - 14C]NAD存在的情况下，通过紫外线照射对产气荚膜梭菌iota毒素的肌动蛋白ADP - 核糖基化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了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 - Gly - X - Tyr - Glu - Val - Leu - Leu - Asn - His - Gly - Ser - Lys，与产气荚膜梭菌iota毒素中从Gly - 363到Lys - 388的区域相对应。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，除了通过与其他ADP - 核糖基转移酶类比得出的Glu - 380外，Glu - 378可能在产气荚膜梭菌iota毒素的活性位点中起关键作用。
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