Structural Studies of Bacterial Luciferase: Subunit Folding and Dimer Assembly in Vivo

新翻译的线性多肽如何折叠成 功能性天然蛋白质是蛋白质研究的中心问题 生物化学今天 拟议研究的目的是 描述了细菌的亚单位 荧光素酶折叠并组装成功能性异二聚体 体内酶 两个亚基luxA和luxB的基因， 在E.杆菌 因此， 可以很容易地制备。 此外，它们的结构可以 易被阿托伐他汀导向的位点特异性改变 诱变 低光强度可以很容易地 测量允许检测突变体中的特定活性 在六个数量级的范围内。 因此，荧光素酶 异源二聚体为研究 控制蛋白质在体内折叠的原则。 在这 将准备亚基的项目突变体， 它们联合收割机结合形成活性酶， 折叠的动力学和物理特性， 组装过程。

How a newly translated linear polypeptide folds into a functional native protein is the central issue in protein biochemistry today. The purpose of the proposed research is to delineate the mechanism by which the subunits of bacterial luciferase fold and assemble into the functional heterodimeric enzyme in vivo. Genes for the two subunits, luxA and luxB, have been cloned and expressed in E. coli. Thus, the subunits can be easily prepared. Moreover, their structures can be readily altered by oligonucleotide-directed site specific mutagenesis. The ease with which low light intensities can be measured permits the detection of specific activites in mutants over a range of six orders of magnitude. Thus, the luciferase heterodimer provides an ideal system for studying the principles which govern protein folding in vivo. In this project mutants of the subunits will be prepared, and the way they combine to form active enzyme will be analyzed through kinetic and physical characterization of the folding and assembly process.