Noguera Martín E, Roman Ernesto A, Rigal Juan B, Cousido-Siah Alexandra, Mitschler André, Podjarny Alberto, Santos Javier
Instituto de Química y Físico-Química Biológicas, Universidad de Buenos Aires, Junín 956, 1113AAD, Buenos Aires, Argentina.
J Biol Inorg Chem. 2015 Jun;20(4):653-64. doi: 10.1007/s00775-015-1251-9. Epub 2015 Apr 2.
Frataxin is an evolutionary conserved protein that participates in iron metabolism. Deficiency of this small protein in humans causes a severe neurodegenerative disease known as Friedreich's ataxia. A number of studies indicate that frataxin binds iron and regulates Fe-S cluster biosynthesis. Previous structural studies showed that metal binding occurs mainly in a region of high density of negative charge. However, a comprehensive characterization of the binding sites is required to gain further insights into the mechanistic details of frataxin function. In this work, we have solved the X-ray crystal structures of a cold-adapted frataxin from a psychrophilic bacterium in the presence of cobalt or europium ions. We have identified a number of metal-binding sites, mainly solvent exposed, several of which had not been observed in previous studies on mesophilic homologues. No major structural changes were detected upon metal binding, although the structures exhibit significant changes in crystallographic B-factors. The analysis of these B-factors, in combination with crystal packing and RMSD among structures, suggests the existence of localized changes in the internal motions. Based on these results, we propose that bacterial frataxins possess binding sites of moderate affinity for a quick capture and transfer of iron to other proteins and for the regulation of Fe-S cluster biosynthesis, modulating interactions with partner proteins.
铁调素是一种参与铁代谢的进化保守蛋白。人类缺乏这种小蛋白会导致一种严重的神经退行性疾病,即弗里德赖希共济失调症。多项研究表明,铁调素能结合铁并调节铁硫簇的生物合成。先前的结构研究表明,金属结合主要发生在高密度负电荷区域。然而,需要对结合位点进行全面表征,以进一步深入了解铁调素功能的机制细节。在这项工作中,我们解析了来自嗜冷细菌的冷适应铁调素在钴离子或铕离子存在下的X射线晶体结构。我们确定了多个金属结合位点,主要暴露于溶剂中,其中一些在先前对嗜温同源物的研究中未被观察到。尽管结构在晶体学B因子上表现出显著变化,但金属结合后未检测到主要的结构变化。对这些B因子的分析,结合晶体堆积和结构间的均方根偏差,表明内部运动存在局部变化。基于这些结果,我们提出细菌铁调素具有中等亲和力的结合位点,用于快速捕获铁并将其转移到其他蛋白质以及调节铁硫簇的生物合成,从而调节与伴侣蛋白的相互作用。