Identification and characterization of a periplasmic trilactone esterase, Cee, revealed unique features of ferric enterobactin acquisition in Campylobacter.
Identification and characterization of a periplasmic trilactone esterase, Cee, revealed unique features of ferric enterobactin acquisition in Campylobacter.
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DOI：
10.1111/mmi.12118
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发表时间：
2013-02
影响因子：
3.6
通讯作者：
Lin J
中科院分区：
文献类型：
--
作者：
Zeng X;Mo Y;Xu F;Lin J
铁摄取是革兰氏阴性菌体内一种高效、保守的铁清除系统。最近，我们在空肠弯曲菌和大肠杆菌中鉴定了两种FeEnt受体(CFRA和CFRB)，这是一种不产生铁载体的肠道人类病原体。在本研究中，通过全基因组测序和比较基因组分析，在空肠弯曲菌中发现了一个独特的Ent三内酯酯酶Cee(Cj1376)。基因组分析和生化分析有力地证明了Cee是空肠弯曲菌中唯一的三内酯酯酶。薄层层析和高效液相色谱分析表明，纯化的CEE对Ent具有较高的水解率。我们还纯化了三个以前从其他细菌(IroE、Irod和Fes)中发现的Cee同源物，并与Cee一起进行了分析，结果表明，Cee、FeS和Irod对Ent的载脂蛋白和铁形式的催化动力学相似，而IroE对Ent的催化效率较低。与细胞质Fes和Irod不同，CEE定位于周质，通过使用CEE特异性抗体的免疫印迹证实。对不同的弯曲菌菌株的遗传操作表明，CEE不仅是CFRB依赖的FeEnt获得所必需的，而且还参与了CFRA依赖的途径。总之，本研究鉴定并鉴定了一种新的周质三内酯酯酶，并提出了一种在弯曲杆菌中获取FeEnt的新模式。
Ferric enterobactin (FeEnt) acquisition is a highly efficient and conserved iron scavenging system in Gram-negative bacteria. Recently, we have characterized two FeEnt receptors (CfrA and CfrB) in Campylobacter jejuni and C. coli, the enteric human pathogens that do not produce any siderophores. In this study, whole genome sequencing and comparative genomic analysis identified a unique Ent trilactone esterase Cee (Cj1376) in C. jejuni. Genomic analysis and biochemical assay strongly suggested that Cee is the sole trilactone esterase in C. jejuni. Thin layer chromatography and HPLC analyses showed high efficiency of the purified Cee to hydrolyze Ent. Three Cee homologs previously characterized from other bacteria (IroE, IroD, and Fes) were also purified and analyzed together with Cee, indicating that Cee, Fes, and IroD displayed similar hydrolysis dynamics for both apo and ferric forms of Ent while IroE catalyzed Ent inefficiently. Unlike cytoplasmic Fes and IroD, Cee is localized in the periplasm as demonstrated by immunoblotting using Cee-specific antibodies. Genetic manipulation of diverse Campylobacter strains demonstrated that Cee is not only essential for CfrB-dependent FeEnt acquisition but also involved in CfrA-dependent pathway. Together, this study identified and characterized a novel periplasmic trilactone esterase and suggested a new model of FeEnt acquisition in Campylobacter.
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DOI：
10.1073/pnas.0737682100
发表时间：
2003-04-01
影响因子：
11.1
作者：
Hantke, K;Nicholson, G;Winkelmann, G
通讯作者：
Winkelmann, G
影响因子：
3.1
作者：
Lin, J;Sahin, O;Zhang, OJ
通讯作者：
Zhang, OJ
影响因子：
4.9
作者：
Lin, J;Michel, LO;Zhang, QJ
通讯作者：
Zhang, QJ
影响因子：
3.1
作者：
Hofreuter, Dirk;Tsai, Jennifer;Galan, Jorge E.
通讯作者：
Galan, Jorge E.
肠杆菌素水解酶 IroE 的结构特征
影响因子：
2.9
作者：
Larsen, N. A.;Lin, H.;Walsh, C. T.
通讯作者：
Walsh, C. T.