Engineering an Fe SOD Active Site into a Host Protein

这项用于探索性研究的小额赠款得到了离子颗粒、生物无机和金属有机化学计划的支持，该计划旨在开发一种新的合成功能金属蛋白的路线。耶鲁大学化学系的John Caradonna博士将使用一种组合方法来构建在已知结构的宿主蛋白质中具有活性金属位点的蛋白质。有效地，反应启动了对满足活性部位几何形状的骨架位置星座的搜索，而不影响宿主蛋白质的三级结构。构建的突变蛋白将进行酶活性测试。最初的实验将试图合成超氧化物歧化酶的铁位置，因此成功的结果可能会导致一个强大的工具，以减少由超氧化物造成的组织损伤。一个直接的结果将是能够测试二级蛋白质拓扑对金属蛋白质化学的影响。含有金属的酶的活性部位由结合到以一定模式折叠的蛋白质上的金属组成。通过设计一种在不破坏折叠模式的情况下添加金属的方法，这项研究可能会导致一种合成具有功能的金属酶的方法，从而准备好用于生物研究和应用。在此过程中，将研究蛋白质结构对金属酶活性的影响。

This Small Grant for Exploratory Research is supported by the Inogranic, Bioinorganic, and Organometallic Chemistry Program for development of a novel synthetic route to functional metalloproteins. Dr. John Caradonna of the Department of Chemistry, Yale University, will use a combinatorial approach to construct proteins with active metal sites inside host proteins of known structure. Effectively, reaction initiates a search for the constellation of backbone positions that satisfies the geometry of the active site without affecting the host protein's tertiary structure. The mutant proteins constructed would be tested for enzymatic activity. The initial experiments will attempt to synthesize the iron site of superoxide dismutase, so a successful outcome could lead to a powerful tool in reducing tissue damage resulting from superoxide. One immediate result will be the ability to test the effect of secondary protein topologies on the chemistry of metalloproteins. The active sites of metal-containing enzymes consist of metals bound to proteins folded in a certain pattern. By devising a means of adding the metal without disrupting the folding pattern, this research could lead to a method of synthesizing metalloenzymes that are functional, and thus ready for use in biological investigations and applications. In the process, the effect of protein structure on activity of metalloenzymes will be studied.