Conservation of an ATP-binding domain among RecA proteins from Proteus vulgaris, Erwinia carotovora, Shigella flexneri, and Escherichia coli K-12 and B/r.
Conservation of an ATP-binding domain among RecA proteins from Proteus vulgaris, Erwinia carotovora, Shigella flexneri, and Escherichia coli K-12 and B/r.
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来自普通变形杆菌、胡萝卜软腐欧文氏菌、福氏志贺氏菌以及大肠杆菌 K-12 和 B/r 的 RecA 蛋白中 ATP 结合域的保守性。
DOI：
10.1128/jb.170.6.2427-2432.1988
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发表时间：
1988
影响因子：
3.2
通讯作者：
McEntee,K
中科院分区：
文献类型：
--
作者：
Knight,KL;Hess,RM;McEntee,K
从普通变形杆菌、胡萝卜软腐欧文氏菌、福氏志贺菌和大肠杆菌B/r中克隆的RecA基因编码的纯化RecA蛋白与从大肠杆菌中克隆的RecA蛋白进行了比较。coli K-12。每种蛋白质在单链DNA存在下水解ATP，并且每种蛋白质都用光亲和ATP类似物8-叠氮腺苷5 '-三磷酸（8 N3 ATP）共价修饰。四个异源RecA蛋白的二维胰蛋白酶图谱显示这些细菌属之间具有相当大的结构保守性。此外，当用胰蛋白酶消化[α-32 P] 8 N3 ATP-修饰的蛋白质并通过高效液相色谱分析时，在每种大肠杆菌中检测到单一放射性峰，并且这些肽与大肠杆菌的胰蛋白酶肽T31相同地洗脱。coliK-12 RecA蛋白，该蛋白是8 N3 ATP光标记的唯一位点。每个异源recA基因与来自大肠杆菌ATP结合结构域序列的寡核苷酸探针杂交。coliK-12基因。这些最后的结果表明，RecA蛋白的ATP结合结构域已经高度保守超过10（7）年。
The purified RecA proteins encoded by the cloned genes from Proteus vulgaris, Erwinia carotovora, Shigella flexneri, and Escherichia coli B/r were compared with the RecA protein from E. coli K-12. Each of the proteins hydrolyzed ATP in the presence of single-stranded DNA, and each was covalently modified with the photoaffinity ATP analog 8-azidoadenosine 5'-triphosphate (8N3ATP). Two-dimensional tryptic maps of the four heterologous RecA proteins demonstrated considerable structural conservation among these bacterial genera. Moreover, when the [alpha-32P]8N3ATP-modified proteins were digested with trypsin and analyzed by high-performance liquid chromatography, a single peak of radioactivity was detected in each of the digests and these peptides eluted identically with the tryptic peptide T31 of the E. coli K-12 RecA protein, which was the unique site of 8N3ATP photolabeling. Each of the heterologous recA genes hybridized to oligonucleotide probes derived from the ATP-binding domain sequence of the E. coli K-12 gene. These last results demonstrate that the ATP-binding domain of the RecA protein has been strongly conserved for greater than 10(7) years.