STRUCTURE, MECHANISM AND REGULATION OF GLYCEROL KINASE

对E. 大肠杆菌磷酸转移酶系统，甘油激酶和因子IIIglc。 长期目标是确定三维结构 单独的蛋白质和复合在一起的蛋白质在生物学上 相关形式。 结构研究将为 甘油激酶的酶学机理研究， 活性变构调节的结构基础， 蛋白质-蛋白质复合物的性质， 磷酸化依赖的信号转导复合物。 已经获得了四种特别高质量的晶体形式， 甘油激酶在底物、产物和 变构效应物 得到了两种有用的晶型 甘油激酶和因子IIIglc之间的复合物， 已经获得了单独的因子IIIglc的晶体。 标准 多个同晶置换程序，分子 将使用置换和非晶体学对称平均 来解决这些晶体形式的结构。 具体 将要解决的问题是：结构基础是什么 用于甘油的酶活性和速度调节 激酶？活性的不同变构效应物是否调节 以相似或不同的方式来构建结构？的本质是什么 依赖磷酸化的蛋白质间相互作用并 甘油激酶具有与其它甘油激酶相同的结构特征， ATP酶？

Structural studies will be carried out for two proteins of the E. coli phosphotransferase system, glycerol kinase and factor IIIglc. The long term goal is to determine the three-dimensional structure of the proteins alone and complexed together in biologically relevant forms. The structural studies will provide the basis for the investigation of the enzymatic mechanism of glycerol kinase, the structural basis for the allosteric regulation of the activity and the nature of the protein-protein complex, which is a phosphorylation dependent signal transduction complex. Four exceptionally high quality crystal forms have been obtained of glycerol kinase in the presence of substrates, products and allosteric effectors. Two useful crystal forms have been obtained of the complex between glycerol kinase and factor IIIglc, and good crystals have been obtained of factor IIIglc alone. The standard procedures of multiple isomorphous replacement, molecular replacement and noncrystallographic symmetry averaging will be used to solve the structures of these crystal forms. The specific questions which will be addressed are: What is the structural basis for the enzymatic activity and velocity modulation of glycerol kinase? Do different allosteric effectors of the activity modulate the structure in similar or different ways? What is the nature of a phosphorylation dependent protein-protein interaction? Does glycerol kinase have structural features in common with other ATPases?