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Expression, Purification and Characterization of a ZIP Family Transporter from Desulfovibrio vulgaris
The Protein Journal ( IF 1.9 ) Pub Date : 2021-06-08 , DOI: 10.1007/s10930-021-10008-7
Cheng Ma 1 , Caixia Gong 2
Affiliation  

The ZIP family transport zinc ions from the extracellular medium across the plasma membrane or from the intracellular compartments across endomembranes, which play fundamental roles in metal homeostasis and are broadly involved in physiological and pathological processes. Desulfovibrio is the predominant sulphate-reducing bacteria in human colonic microbiota, but also a potential choice for metal bioremediation. while, there are no published studies describing the zinc transporters from Desulfovibrio up to now. In this study, we obtained for the first time a heterologously expressed ZIP homolog from Desulfovibrio vulgaris, termed dvZip. The purified dvZip was reconstituted into proteoliposomes, and confirmed its zinc transport ability in vitro. By combining topology prediction, homology modeling and phylogenetic approaches, we also noticed that dvZip belongs to the GufA and probably have 8 transmembrane α-helical segments (TM 1–8) in which both termini are located on the extracellular, with TM2, 4, 5 and 7 create an inner bundle. We believe that purification and characterization of zinc (probably also cadmium) transporters from Desulfovibrio vulgaris such as dvZip could shed light on understanding of metal homeostasis of Desulfovibrio and provided protein products for future detailed function and structural studies.



中文翻译:

普通脱硫弧菌 ZIP 家族转运蛋白的表达、纯化和表征

ZIP 家族将锌离子从细胞外介质穿过质膜或从细胞内隔室穿过内膜转运,这在金属稳态中发挥着重要作用,并广泛参与生理和病理过程。脱硫弧菌是人类结肠微生物群中主要的硫酸盐还原菌,也是金属生物修复的潜在选择。而到目前为止,还没有发表的研究描述来自脱硫弧菌的锌转运蛋白。在本研究中,我们首次从普通脱硫弧菌中获得了异源表达的 ZIP 同源物,称为 dvZip。纯化后的 dvZip 被重组为蛋白脂质体,并在体外证实了其锌转运能力。通过结合拓扑预测、同源性建模和系统发育方法,我们还注意到 dvZip 属于 GufA,可能有 8 个跨膜 α-螺旋段(TM 1-8),其中两个末端都位于细胞外,TM2、4、 5 和 7 创建一个内包。我们相信,从普通脱硫弧菌(如 dvZip )中纯化和表征锌(可能还有镉)转运蛋白可以阐明对脱硫弧菌的金属稳态的理解,并为未来详细的功能和结构研究提供蛋白质产品。

更新日期:2021-06-08
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