CAREER: The Evolution of Phosphoryl Transfer and Protein Sequence Analysis

9984919 Sanders在该提案中，PI希望通过研究一个古老而多样的磷酸转移酶家族的进化来更深入地了解蛋白质一级结构的信息内容。 他将专注于从嗜热甲烷八叠球菌乙酸激酶的结构的X射线晶体学测定。 本课程将探讨金属离子与核苷酸结合的化学细节、催化的机制以及每个保守氨基酸残基在蛋白质结构和功能中的作用。 从这些分析中，他希望深入了解在糖激酶/肌动蛋白/Hsc 70磷酸转移酶超家族分化过程中发生的底物结合和催化模式的变化。 已被添加到核心蛋白折叠在家庭的各个成员的二级结构的元素的意义将进行调查。 丁酸激酶和外聚磷酸酶的磷酰基转移催化也将通过结构，生物化学和诱变方法的组合进行检查。 海栖热袍菌丁酸激酶是否通过一个或多个共价磷酸酶中间体催化磷酰基转移的检查将进行。 将进一步开发为高水平本科生介绍蛋白质序列分析而设计的课程。 它将包括一个蛋白质序列的解释如何取决于不同序列的比较和蛋白质中氨基酸残基的上下文依赖性化学的理解的检查。 学生将有机会探索计算机生成的蛋白质结构表示，并执行和探测蛋白质序列分析的原则。 将编写和分发可能对其他研究人员和学生有益的教材。

9984919SandersIn this proposal, the PI wishes to achieve a deeper understanding of the information content of the primary structure of a protein by studies of the evolution of an ancient and diverse family of phosphotransferases. He will concentrate on the X-ray crystallographic determination of the structure of acetate kinase from Methanosarcina thermophila. The chemical details of metal-ion and nucleotide binding, the mechanism of catalysis and the roles of each conserved amino-acid residue in protein structure and function will be explored. From these analyses, he wishes to gain insight into the changes in the modes of substrate binding and catalysis that occurred during the divergence of the sugar kinase/actin/Hsc70 super-family of phosphotransferases. The significance of the elements of secondary structure that have been added to the core protein fold in various members of the family will be investigated. Catalysis of phosphoryl transfer by butyrate kinases and an exopolyphosphatase will also be examined through a combination of structural, biochemical, and mutagenesis approaches. An examination of whether the Thermotoga maritima butyrate kinase catalyzes phosphoryl transfer through one or more covalent phosphoenzyme intermediates will be undertaken. A course designed for the introduction of upper level undergraduates to protein sequence analysis will be developed further. It will encompass an examination of how the interpretation of a protein sequence depends upon both comparison of different sequences and an understanding of the context-dependent chemistry of the amino-acid residues in proteins. The students will have the opportunity to explore computer-generated representations of protein structures and to perform and probe the principles of protein-sequence analyses. Course materials that may be beneficial to other researchers and students will be constructed and disseminated.