A putative siderophore-interacting protein from the marine bacterium Shewanella frigidimarina NCIMB 400: cloning, expression, purification, crystallization and X-ray diffraction analysis.
A putative siderophore-interacting protein from the marine bacterium Shewanella frigidimarina NCIMB 400: cloning, expression, purification, crystallization and X-ray diffraction analysis.
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DOI：
10.1107/s2053230x16011419
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发表时间：
2016-09
期刊：
影响因子：
--
通讯作者：
Moe E
中科院分区：
文献类型：
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作者：
Trindade IB;Fonseca BM;Matias PM;Louro RO;Moe E
成功克隆了海洋细菌S. frigidimarina中铁载体相互作用蛋白的编码基因，并对其进行了表达和纯化。优化的晶体衍射到1.35 Å分辨率和初步的晶体学分析在结构确定和增加对尚不清楚的铁获取分子机制的了解方面是有希望的。铁载体结合蛋白（SIPs）在多种生物的铁获取中起着关键作用。在海洋细菌冷希瓦氏菌NCIMB 400的基因组中，标记为SFRI_RS12295的基因编码来自该家族的蛋白质。本文报道了该蛋白的克隆、表达、纯化和结晶，并对其进行了初步的x射线晶体学分析，分辨率为1.35 Å。SIP晶体属于单斜空间群P21，单位胞参数a = 48.04, b = 78.31, c = 67.71 Å， α = 90, β = 99.94, γ = 90°，预测每个不对称单元含有2个分子。通过分子替换确定结构，并使用先前确定的具有~ 30%序列同一性的~ 2 Å分辨率SIP结构作为模板，目前正在进行中。
The gene encoding a putative siderophore-interacting protein from the marine bacterium S. frigidimarina was successfully cloned, followed by expression and purification of the gene product. Optimized crystals diffracted to 1.35 Å resolution and preliminary crystallographic analysis is promising with respect to structure determination and increased insight into the poorly understood molecular mechanisms underlying iron acquisition. Siderophore-binding proteins (SIPs) perform a key role in iron acquisition in multiple organisms. In the genome of the marine bacterium Shewanella frigidimarina NCIMB 400, the gene tagged as SFRI_RS12295 encodes a protein from this family. Here, the cloning, expression, purification and crystallization of this protein are reported, together with its preliminary X-ray crystallographic analysis to 1.35 Å resolution. The SIP crystals belonged to the monoclinic space group P21, with unit-cell parameters a = 48.04, b = 78.31, c = 67.71 Å, α = 90, β = 99.94, γ = 90°, and are predicted to contain two molecules per asymmetric unit. Structure determination by molecular replacement and the use of previously determined ∼2 Å resolution SIP structures with ∼30% sequence identity as templates are ongoing.