Zinc coordination is essential for the function and activity of the type II secretion ATPase EpsE.
Zinc coordination is essential for the function and activity of the type II secretion ATPase EpsE.
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DOI：
10.1002/mbo3.376
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发表时间：
2016-10
期刊：
影响因子：
3.4
通讯作者：
Sandkvist, Maria
中科院分区：
文献类型：
--
作者：
Rule, Chelsea S.;Patrick, Marcella;Camberg, Jodi L.;Maricic, Natalie;Hol, Wim G.;Sandkvist, Maria
霍乱弧菌中的II型分泌系统Eps促进霍乱毒素和几种水解酶的细胞外转运，并且是许多革兰氏阴性病原体中的主要毒力系统，其在结构上与IV型菌毛系统相关。细胞质ATP酶EpsE通过ATP水解为胞外蛋白分泌提供能量。EpsE含有一个独特的金属结合结构域，通过四半胱氨酸基序（CXXCX 29 CXXC）与锌配位，该基序也存在于IV型菌毛组装中，但不存在于收缩ATP酶中。缺失整个结构域或取代任何与锌配位的半胱氨酸残基完全消除EpsE缺陷型菌株中的分泌，并且在野生型EpsE存在下对分泌具有显性负效应。与体内数据一致，从纯化的EpsE六聚体中化学消耗锌导致体外ATP酶活性丧失。相反，将两个双半胱氨酸之间的残基与来自铜绿假单胞菌的同源ATP酶XcpR的残基交换对EpsE没有显著影响。这些结果表明，尽管金属结合结构域中的单个残基通常是可互换的，但锌配位对于EpsE的活性和功能是必不可少的。
The type II secretion system Eps in Vibrio cholerae promotes the extracellular transport of cholera toxin and several hydrolytic enzymes and is a major virulence system in many Gram‐negative pathogens which is structurally related to the type IV pilus system. The cytoplasmic ATPase EpsE provides the energy for exoprotein secretion through ATP hydrolysis. EpsE contains a unique metal‐binding domain that coordinates zinc through a tetracysteine motif (CXXCX29CXXC), which is also present in type IV pilus assembly but not retraction ATPases. Deletion of the entire domain or substitution of any of the cysteine residues that coordinate zinc completely abrogates secretion in an EpsE‐deficient strain and has a dominant negative effect on secretion in the presence of wild‐type EpsE. Consistent with the in vivo data, chemical depletion of zinc from purified EpsE hexamers results in loss of in vitro ATPase activity. In contrast, exchanging the residues between the two dicysteines with those from the homologous ATPase XcpR from Pseudomonas aeruginosa does not have a significant impact on EpsE. These results indicate that, although the individual residues in the metal‐binding domain are generally interchangeable, zinc coordination is essential for the activity and function of EpsE.
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影响因子：
16
作者：
Reindl, Sophia;Ghosh, Abhrajyoti;Williams, Gareth J.;Lassak, Kerstin;Neiner, Tomasz;Henche, Anna-Lena;Albers, Sonja-Verena;Tainer, John A.
通讯作者：
Tainer, John A.
影响因子：
3.6
作者：
Gray MD;Bagdasarian M;Hol WG;Sandkvist M
通讯作者：
Sandkvist M
影响因子：
5.6
作者：
Misic, Ana M.;Satyshur, Kenneth A.;Forest, Katrina T.
通讯作者：
Forest, Katrina T.
DOI：
10.1073/pnas.051436598
发表时间：
2001-02-27
影响因子：
11.1
作者：
Planet, PJ;Kachlany, SC;Figurski, DH
通讯作者：
Figurski, DH
影响因子：
2.8
作者：
Chiang, Poney;Sampaleanu, Liliana M.;Burrows, Lori L.
通讯作者：
Burrows, Lori L.