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A unique ferrous iron binding mode is associated with large conformational changes for the transport protein FpvC of Pseudomonas aeruginosa.
The FEBS Journal ( IF 5.5 ) Pub Date : 2019-07-26 , DOI: 10.1111/febs.15004
Armelle Vigouroux 1 , Magali Aumont-Nicaise 1 , Alain Boussac 1 , Loïc Marty 1 , Léa Lo Bello 1 , Pierre Legrand 2 , Karl Brillet 3 , Isabelle J Schalk 3 , Solange Moréra 1
Affiliation  

Pseudomonas aeruginosa secretes pyoverdine, a major siderophore to get access to iron, an essential nutrient. Pyoverdine scavenges ferric iron in the bacterial environment with the resulting complex internalized by bacteria. Releasing of iron from pyoverdine in the periplasm involves an iron reduction by an inner membrane reductase and two solute-binding proteins (SBPs) FpvC and FpvF in association with their ABC transporter. FpvC and FpvF belong to two different subgroups of SBPs within the structural cluster A: FpvC and FpvF were proposed to be a metal-binding protein and a ferrisiderophore-binding protein respectively. Here, we report the redox state and the binding mode of iron to FpvC. We first solved the crystal structure of FpvC bound to a fortuitous Ni2+ by single anomalous dispersion method. Using a different protein purification strategy, we determined the structure of FpvC with manganese and iron, which binds to FpvC in a ferrous state as demonstrated by electron paramagnetic resonance. FpvC is the first example of a hexahistidine metal site among SBPs in which the Fe2+ redox state is stabilized under aerobic conditions. Using biophysics methods, we showed that FpvC reversibly bind to a broad range of divalent ions. The structure of a mutant mimicking the apo FpvC reveals a protein in an open state with large conformational changes when compared with the metal-bound FpvC. These results highlight that the canonical metal site in FpvC is distinct from those yet described in SBPs and they provide new insights into the mechanism of PVD-Fe dissociation in P. aeruginosa.

中文翻译:

铜绿假单胞菌转运蛋白FpvC的独特构象变化与独特的亚铁结合模式有关。

铜绿假单胞菌分泌pyoverdine,这是一种主要的铁载体,可接触铁(一种基本营养素)。Pyoverdine在细菌环境中清除了三价铁,并且所产生的复合物被细菌内在化。从周质中的嘧啶定释放铁涉及通过内膜还原酶和两个溶质结合蛋白(SBP)FpvC和FpvF及其ABC转运蛋白来还原铁。FpvC和FpvF属于结构簇A内SBP的两个不同亚组:FpvC和FpvF分别被认为是金属结合蛋白和亚铁金属结合蛋白。在这里,我们报告氧化还原状态和铁对FpvC的绑定模式。我们首先通过单一异常分散方法解决了与偶然Ni2 +结合的FpvC的晶体结构。使用不同的蛋白质纯化策略,我们通过锰和铁确定了FpvC的结构,该结构以亚铁态与FpvC结合,如电子顺磁共振所证实的那样。FpvC是SBP中六组氨酸金属位点的第一个例子,其中Fe2 +氧化还原态在好氧条件下稳定。使用生物物理学方法,我们表明FpvC可逆地结合到广泛的二价离子上。与金属结合的FpvC相比,模仿apo FpvC的突变体的结构揭示了一种处于开放状态的蛋白质,其构象变化很大。这些结果表明,FpvC中的典型金属位点与SBP中尚未描述的位点不同,它们为铜绿假单胞菌中PVD-Fe解离的机理提供了新的见解。如电子顺磁共振所证实,其以亚铁状态结合至FpvC。FpvC是SBP中六组氨酸金属位点的第一个例子,其中Fe2 +氧化还原态在好氧条件下稳定。使用生物物理学方法,我们表明FpvC可逆地结合到广泛的二价离子上。与金属结合的FpvC相比,模仿apo FpvC的突变体的结构揭示了一种处于开放状态的蛋白质,其构象变化很大。这些结果表明,FpvC中的典型金属位点与SBP中尚未描述的位点截然不同,它们为铜绿假单胞菌中PVD-Fe解离的机理提供了新的见解。如电子顺磁共振所证实,其以亚铁状态与FpvC结合。FpvC是SBP中六组氨酸金属位点的第一个例子,其中Fe2 +氧化还原态在好氧条件下稳定。使用生物物理学方法,我们表明FpvC可逆地结合到广泛的二价离子上。与金属结合的FpvC相比,模仿apo FpvC的突变体的结构揭示了一种处于开放状态的蛋白质,其构象变化很大。这些结果表明,FpvC中的典型金属位点与SBP中尚未描述的位点截然不同,它们为铜绿假单胞菌中PVD-Fe解离的机理提供了新的见解。使用生物物理学方法,我们表明FpvC可逆地结合到广泛的二价离子上。与金属结合的FpvC相比,模仿apo FpvC的突变体的结构揭示了一种处于开放状态的蛋白,其构象变化很大。这些结果表明,FpvC中的典型金属位点与SBP中尚未描述的位点截然不同,它们为铜绿假单胞菌中PVD-Fe解离的机理提供了新的见解。使用生物物理学方法,我们表明FpvC可逆地结合到广泛的二价离子上。与金属结合的FpvC相比,模仿apo FpvC的突变体的结构揭示了一种处于开放状态的蛋白质,其构象变化很大。这些结果表明,FpvC中的典型金属位点与SBP中尚未描述的位点截然不同,它们为铜绿假单胞菌中PVD-Fe解离的机理提供了新的见解。
更新日期:2020-01-21
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