STRUCTURE, MECHANISM AND REGULATION OF GLYCEROL KINASE

将对E. Coli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 单独的蛋白质和在生物上复合在一起的蛋白质 相关表格。结构研究将为 甘油激酶的酶作用机制的研究 活性和变构调节的结构基础 蛋白质-蛋白质复合体的性质，这是一种 磷酸化依赖的信号转导复合体。 已经获得了四种极高质量的晶体形式 存在底物、产物和甘油激酶 变构效应器。得到了两种有用的晶型 甘油激酶和因子IIIglc之间的复合体，以及Good 单单得到了因子IIIglc的晶体。标准 多重同象取代的过程，分子 将使用替换和非晶体对称性平均 来解开这些晶型的结构。具体的 将解决的问题是：结构基础是什么 甘油的酶活性和速度调节 激活剂？不同的变构活性效应剂会不会进行调节？ 这种结构是相似的还是不同的？其本质是什么？ 依赖于磷酸化的蛋白质-蛋白质相互作用？会吗？ 甘油激酶具有与其他 ATPase？

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?