Department of Chemistry, Faculty of Science and Technology, The Norwegian Structural Biology Centre (NorStruct), UiT the Arctic University of Norway, 9037, Tromsø, Norway.
Biometals. 2020 Oct;33(4-5):169-185. doi: 10.1007/s10534-020-00240-6. Epub 2020 Jul 9.
Iron is an essential nutrient for bacteria, however its propensity to form toxic hydroxyl radicals at high intracellular concentrations, requires its acquisition to be tightly regulated. Ferric uptake regulator (Fur) is a metal-dependent DNA-binding protein that acts as a transcriptional regulator in maintaining iron metabolism in bacteria and is a highly interesting target in the design of new antibacterial drugs. Fur mutants have been shown to exhibit decreased virulence in infection models. The protein interacts specifically with DNA at binding sites designated as 'Fur boxes'. In the present study, we have investigated the interaction between Fur from the fish pathogen Aliivibrio salmonicida (AsFur) and its target DNA using a combination of biochemical and in silico methods. A series of target DNA oligomers were designed based on analyses of Fur boxes from other species, and affinities assessed using electrophoretic mobility shift assay. Binding strengths were interpreted in the context of homology models of AsFur to gain molecular-level insight into binding specificity.
铁是细菌必需的营养物质，然而，由于其在细胞内高浓度下容易形成有毒的羟基自由基，因此需要严格控制其摄取。亚铁摄取调节蛋白（Fur）是一种金属依赖性 DNA 结合蛋白，作为转录调节剂，在维持细菌的铁代谢中发挥作用，是设计新型抗菌药物的极具吸引力的靶标。研究表明，Fur 突变体在感染模型中表现出毒力降低。该蛋白在称为“Fur 盒”的结合位点上与 DNA 特异性相互作用。在本研究中，我们结合生化和计算方法研究了鱼类病原体杀鲑气单胞菌（AsFur）的 Fur 与其靶 DNA 之间的相互作用。根据来自其他物种的 Fur 盒分析设计了一系列靶 DNA 寡核苷酸，并使用电泳迁移率变动分析评估了亲和力。在 AsFur 的同源模型背景下解释结合强度，以获得分子水平的结合特异性见解。
