Engineering an Fe SOD Active Site into a Host Protein

这项探索性研究小额赠款由无机、生物无机和有机金属化学计划提供支持，用于开发一种新的合成功能性金属蛋白的途径。耶鲁大学化学系的John Caradonna博士将使用组合方法在已知结构的宿主蛋白质中构建具有活性金属位点的蛋白质。有效地，反应启动了在不影响宿主蛋白三级结构的情况下，满足活性位点几何形状的主链位置星座的搜索。构建的突变蛋白将进行酶活性测试。最初的实验将尝试合成超氧化物歧化酶的铁位点，因此一个成功的结果可能会成为减少超氧化物引起的组织损伤的有力工具。一个直接的结果将是测试二级蛋白拓扑结构对金属蛋白化学的影响。含金属酶的活性位点是由以特定模式折叠的与蛋白质结合的金属组成的。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功能性金属酶的方法，从而为生物研究和应用做好准备。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.