Aminoglycoside multiacetylating activity of the enhanced intracellular survival protein from Mycobacterium smegmatis and its inhibition.
Aminoglycoside multiacetylating activity of the enhanced intracellular survival protein from Mycobacterium smegmatis and its inhibition.
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DOI：
10.1021/bi3004473
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发表时间：
2012-06-19
期刊：
影响因子：
2.9
通讯作者：
Garneau-Tsodikova S
中科院分区：
文献类型：
--
作者：
Chen W;Green KD;Tsodikov OV;Garneau-Tsodikova S
增强的细胞内存活（Eis）蛋白改善了巨噬细胞中的耻垢分枝杆菌（Msm）存活，并在大量结核分枝杆菌（Mtb）临床分离株中充当负责卡那霉素A抗性（广泛耐药（XDR）结核病的标志）的乙酰转移酶。我们最近证明了来自Mtb的Eis（Eis_Mtb）有效地多乙酰化多种氨基糖苷类（AG）抗生素。在这里，为了深入了解Eis对AG多乙酰化的底物选择性的起源，我们分析了Eis_Msm对AG乙酰化的影响，研究了其抑制作用，并将这些功能与Eis_Mtb的功能进行了比较。尽管对于几种AG，Eis_Msm和Eis_Mtb的多乙酰化性质相似，但存在三个主要差异：（i）Eis_Msm使安普霉素二乙酰化，安普霉素是一种构象受限的AG，Eis_Mtb不能对其进行修饰，（ii）Eis_Msm使巴龙霉素三乙酰化，巴龙霉素只能被Eis_Mtb二乙酰化，和（iii）Eis_Msm仅使潮霉素单乙酰化，一种结构独特的AG，被Eis_Mtb二乙酰化。几个非保守的氨基酸残基内衬的AG结合口袋的Eis可能是负责这些差异之间的两个Eis同源物。具体地说，我们提出，因为AG结合口袋的Eis_Msm比Eis_Mtb更开放，它容纳安普霉素乙酰化在Eis_Msm，但不是在Eis_Mtb。我们还证明了我们最近发现的Eis_Mtb抑制剂可以抑制Eis_Msm活性。这些观察结果有助于确定Eis同源物中底物偏好的结构起源，并表明Eis_Mtb抑制剂可用于对抗所有致病性分枝杆菌，以克服由Eis上调引起的AG抗性。
The enhanced intracellular survival (Eis) protein improves Mycobacterium smegmatis (Msm) survival in macrophages and functions as the acetyltransferase responsible for kanamycin A resistance, a hallmark of extensively drug-resistant (XDR) tuberculosis, in a large number of Mycobacterium tuberculosis (Mtb) clinical isolates. We recently demonstrated that Eis from Mtb (Eis_Mtb) efficiently multi-acetylates a variety of aminoglycoside (AG) antibiotics. Here, to gain insight into the origin of substrate selectivity of AG multi-acetylation by Eis, we analyzed AG acetylation by Eis_Msm, investigated its inhibition, and compared these functions to those of Eis_Mtb. Even though for several AGs the multi-acetylation properties of Eis_Msm and Eis_Mtb are similar, there are three major differences: (i) Eis_Msm di-acetylates apramycin, a conformationally constrained AG, which Eis_Mtb cannot modify, (ii) Eis_Msm tri-acetylates paromomycin, which can be only di-acetylated by Eis_Mtb, and (iii) Eis_Msm only mono-acetylates hygromycin, a structurally unique AG that is di-acetylated by Eis_Mtb. Several non-conserved amino acid residues lining the AG-binding pocket of Eis are likely responsible for these differences between the two Eis homologs. Specifically, we propose that because the AG-binding pocket of Eis_Msm is more open than that of Eis_Mtb, it accommodates apramycin for acetylation in Eis_Msm, but not in Eis_Mtb. We also demonstrate that inhibitors of Eis_Mtb that we recently discovered can inhibit Eis_Msm activity. These observations help define the structural origins of substrate preference among Eis homologs and suggest that Eis_Mtb inhibitors may be applied against all pathogenic mycobacteria to overcome AG resistance caused by Eis up-regulation.
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影响因子：
2.9
作者：
Magalhaes, MLB;Blanchard, JS
通讯作者：
Blanchard, JS
影响因子：
2.9
作者：
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DOI：
10.1073/pnas.0907925106
发表时间：
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影响因子：
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影响因子：
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作者：
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作者：
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