CRYSTALLOGRAPHIC STUDIES OF IRE BINDING PROTEIN

说明(改编自应用程序) 本应用程序的目标是研究铁响应的相互作用 蛋白质及其RNA结合部位，使用X射线结晶学。这些 相互作用在铁的运输和调节中发挥核心作用 动态平衡。铁的运输和代谢疾病是最常见的疾病之一 造成人类痛苦和死亡的普遍原因。更好地理解 调节铁的吸收和利用将是重要的#年。 开发铁缺乏和铁超载的治疗方法。铁监管 蛋白质(IRP)对其他蛋白质的水平进行翻译控制 参与铁的新陈代谢。已知的两个IRP是IRP-1和IRP-2。时铁 水平低，这些蛋白质识别特定的茎/环结构，称为 铁反应元件(IRES)，存在于靶mRNAs中。为 在低铁水平下调的蛋白质中，IRE通常被发现 在消息的5‘非翻译区和IRP与IRE的结合中 禁止翻译消息。对于需要在下面表达的蛋白质 在低铁条件下水平较高，IRE通常出现在3‘ 非翻译区，其中IRP结合稳定转录本，导致 蛋白质的表达增加。使用X射线结晶学，我们将 追求IRP-2单独的结构以及与两个已知类别的复杂结构 Irm RNA结合位点数。我们将继续研究IRP-2的晶体结构 具有C凸起型IRE的复合体，以及具有铁蛋白环/凸起型的复合体 愤怒。在IRP-2结构完成后，进一步的结构功能 研究工作将会展开。这些研究将使用定点突变， 生化特性和结构测定有待进一步探索 IRP-2/IRE的结构和功能。这些研究提供的更大洞察力 深入研究IRPS与IRES的具体互动可能会让我们更近一步 朝着铁代谢疾病的治疗方法的发展。

DESCRIPTION (adapted from the application) The goal of this application is to study the interactions of iron response proteins with their RNA binding sites, using X-ray crystallography. These interactions play a central role in the regulation of iron transport and homeostasis. Diseases of iron transport and metabolism are among the most prevalent causes of human suffering and mortality. A greater understanding of the regulation of iron absorption and utilization will be important in developing treatments for iron deficiency and iron overload. Iron regulatory proteins (IRPs) exert translational control over the levels of other proteins involved in iron metabolism. Two IRPs are known, IRP-1 and IRP-2. When iron levels are low, these proteins recognize specific stem/loop structures known as iron response elements (IREs), which are present in the target mRNAs. For proteins whose levels are downregulated in low iron, the IRE is typically found in the 5' untranslated region of the message and binding of the IRP to the IRE inhibits translation of the message. For proteins that need to be expressed at higher levels under low iron conditions, the IRE is generally found in the 3' untranslated region, where IRP binding stabilizes the transcript, resulting in increased expression of the protein. Using X-ray crystallography, we will pursue the structure of IRP-2 alone and in complex with the two known classes of IRE mRNA binding sites. We will pursue crystal structures of IRP-2 in complex with a C bulge type IRE, and in complex with a ferritin loop/bulge type IRE. Upon the completion of IRP-2 structures, further structure function studies will be initiated. These studies will use site directed mutagenesis, biochemical characterization and structure determination to further explore IRP-2/IRE structure and function. The greater insight provided by these studies into the specific interactions of IRPs with IREs may lead us a step closer towards the development of treatments for disease of iron metabolism.