Vibrio cholerae FeoB hydrolyzes ATP and GTP in vitro in the absence of stimulatory factors,Metallomics

当前位置： X-MOL 学术 › Metallomics › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

Vibrio cholerae FeoB hydrolyzes ATP and GTP in vitro in the absence of stimulatory factors
Metallomics ( IF 2.9 ) Pub Date : 2020-11-4 , DOI: 10.1039/d0mt00195c
Camilo Gómez-Garzón ₁ , Shelley M Payne

Affiliation

Feo is the most widely conserved system for ferrous iron transport in prokaryotes, and it is important for virulence in some pathogens. However, its mechanism of iron transport is not fully understood. In this study, we used full-length Vibrio cholerae FeoB (VcFeoB) as a model system to study whether its enzymatic activity is affected by regulatory factors commonly associated with FeoB proteins from other species or with G-proteins that have homology to FeoB. VcFeoB showed a higher rate of hydrolysis of both ATP and GTP than its N-terminal domain alone; likewise, ions such as K⁺ and Fe²⁺ did not modulate its nucleotide hydrolysis. We also showed that the three V. cholerae Feo proteins (FeoA, FeoB, and FeoC) work in a 1 : 1 : 1 molar ratio in vivo. Although both FeoA and FeoC are required for Feo-mediated iron transport, neither of these proteins affected the VcFeoB NTPase rate. These results are consistent with an active transport mechanism independent of stimulatory factors and highlight the importance of using full-length FeoB proteins as a reliable proxy to study Feo-mediated iron transport in vitro.

中文翻译：

在没有刺激因子的情况下，霍乱弧菌 FeoB 在体外水解 ATP 和 GTP

Feo 是原核生物中亚铁转运最保守的系统，它对某些病原体的毒力很重要。然而，其铁转运机制尚不完全清楚。在这项研究中，我们使用全长霍乱弧菌FeoB (VcFeoB) 作为模型系统来研究其酶活性是否受到通常与其他物种的 FeoB 蛋白或与 FeoB 同源的 G 蛋白相关的调节因子的影响。VcFeoB 的 ATP 和 GTP 水解速率均高于单独的 N 端结构域；同样，K ⁺和 Fe ^{2+ 等离子}不会调节其核苷酸水解。我们还表明，三种霍乱弧菌Feo 蛋白（FeoA、FeoB 和 FeoC）在体内以 1:1:1 的摩尔比起作用。尽管 FeoA 和 FeoC 都是 Feo 介导的铁转运所必需的，但这两种蛋白质都不影响 VcFeoB NTPase 速率。这些结果与独立于刺激因素的主动转运机制一致，并强调了使用全长 FeoB 蛋白作为研究 Feo 介导的体外铁转运的可靠代理的重要性。

更新日期：2021-01-06

点击分享查看原文

点击收藏

阅读更多本刊最新论文本刊介绍/投稿指南11