CRYSTALLOGRAPHIC STUDIES OF IRE BINDING PROTEIN

描述（改编自应用程序） 本申请的目的是研究铁反应的相互作用 蛋白质与它们的RNA结合位点，使用X射线晶体学。这些 相互作用在铁转运的调节中起核心作用， 体内平衡铁转运和代谢疾病是最常见的疾病之一。 人类痛苦和死亡的普遍原因。更好地了解 铁吸收和利用的调节将是重要的， 开发铁缺乏和铁过量的治疗方法。铁调节 蛋白质（IRP）对其他蛋白质的水平进行翻译控制 参与铁代谢。已知有两种IRP，IRP-1和IRP-2。当铁 水平低，这些蛋白质识别特定的茎/环结构，称为 铁反应元件（IRE），其存在于靶mRNA中。为 在低铁水平下水平下调的蛋白质中，IRE通常被发现 在信息的5'非翻译区和IRP与IRE的结合 禁止消息的翻译。对于那些需要表达的蛋白质， 在低铁条件下，IRE水平较高，通常存在于3'端。 非翻译区，其中IRP结合稳定转录物，导致 增加蛋白质的表达。使用X射线晶体学，我们将 单独研究IRP-2的结构，并与两个已知的类进行复合 IRE mRNA结合位点。我们将继续研究IRP-2的晶体结构， 与C凸起型IRE复合，并与铁蛋白环/凸起型复合 爱尔兰共和国IRP-2结构完成后， 研究将启动。这些研究将使用定点诱变， 生化表征和结构测定，以进一步探索 IRP-2/IRE结构和功能。这些研究提供的更深刻的见解 IRP与IRE的特定相互作用可能会使我们更接近一步 对铁代谢疾病的治疗的发展。

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.