REGULATION OF POLYSACCHARIDE SYNTHESIS

多糖合成的调控

• 批准号: 2062001
• 负责人: JACK PREISS
• 金额: $ 27.61万
• 依托单位: MICHIGAN STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1985
• 资助国家: 美国
• 起止时间: 1985-05-01 至 1996-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2062001
• 关键词: Enterobacteriaceae; Escherichia coli; Escherichia coli k12; Rhodospirillum; Salmonella typhimurium; affinity chromatography; affinity labeling; allosteric site; bacterial genetics; carbohydrate biosynthesis; carbon; enzyme mechanism; enzyme structure; gel electrophoresis; gel filtration chromatography; gene induction /repression; genetic manipulation; genetic mapping; genetic strain; genetic transcription; genetic translation; glycogen; glycogen synthase; glycosyltransferase; molecular cloning; mutant; nucleic acid sequence; other phosphotransferase; photosynthetic bacteria; protein sequence; radionuclides; radiotracer; structural genes

Studies on the bacterial glycogen biosynthetic enzymes will be continued with respect to elucidating their structure and to relate structure with function and specificity of binding at various ligand binding sites (e.g., inhibitor, activator and catalytic sites). These studies will be centered on the enzyme, ADPglucose pyrophosphorylase. A study of kinetically altered ADPglucose pyrophosphorylases from E. coli glycogen mutants is initiated to determine the relationship of allosteric function with the primary structure of the ADPglucose pyrophosphorylase. The cloning of the structural gene for the ADPglucose pyrophosphorylase of the E. coli allosteric mutant, 618, and the nucleotide sequence studies of it will enable us to determine the amino acid substitution and give us some insight in the study of structure and function of the allosteric site(s). Other E. coli ADPglucose pyrophosphorylase allosteric mutants that have been previously described will also be cloned to provide more information on the structure and function of the allosteric site(s). Cloning of the structural genes of the ADPglucose pyrophosphorylase from other bacteria (e.g., R. rubrum, R. capsulata, R. sphaeroides) are planned and attempts to obtain good expression of the gene are also planned. Nucleotide sequence studies and covalent modification of the various proteins with activator analogues may provide us with useful information of the sequence of the allosteric sites of these ADPglucose pyrophosphorylases. These comparative studies at the protein sequence level will enable us to relate structure with function and activator specificity at these various ligand binding sites. The characterization of E. coli B glycogen synthase and branching enzyme with respect to reaction mechanism and the various amino acids involved in catalysis will be continued. Studies on the cloning of the glycogen biosynthetic structural genes of E. coli K12 will be continued to enable us to determine DNA sequence analysis of the glg (glycogen synthase) structural gene. From this analysis the amino acid sequence will be deduced. The genetic regulation of the biosynthesis of the glycogen biosynthetic enzymes will also be studied and it is hoped that the isolation of the glg genes from both E. coli and Salmonella typhimurium will provide us with some insight in the mode of genetic regulation of expression of the glg genes.

细菌糖原生物合成酶的研究将继续 关于阐明其结构并将结构与 在各种配体结合位点结合的功能和特异性（例如， 抑制剂，激活剂和催化部位）。 这些研究将集中 在酶上，adpglucose焦磷酸化酶。 动力学研究 来自大肠杆菌糖原突变体的adpglucose焦磷酸酶改变了 启动以确定变构功能与 Adpglucose焦磷酸化酶的一级结构。 克隆 大肠杆菌的Adpglucose焦磷酸化酶的结构基因 变构突变体，618和IT的核苷酸序列研究将 使我们能够确定氨基酸取代并给我们一些见识 在对变构位点的结构和功能的研究中。 其他E. 大肠杆菌adpglucose焦磷酸化酶变构突变体 先前描述的也将克隆以提供有关 变构位点的结构和功能。 克隆 来自其他细菌的adpglucose焦磷酸化酶的结构基因 （例如，R。Rubrum，R。Capsulata，R。Sphaeroides），并试图尝试 还计划获得良好的基因表达。 核苷酸序列 各种蛋白质与激活剂的研究和共价修饰 类似物可以为我们提供有关序列的有用信息 这些Adpglucose焦磷酸酶的变构位点。 这些比较 蛋白质序列水平的研究将使我们能够联系结构 在这些各种配体结合处的功能和激活剂特异性 站点。 大肠杆菌B糖原合酶和分支的表征 关于反应机理和各种氨基酸的酶 参与催化将继续进行。 关于克隆的研究 大肠杆菌K12的糖原生物合成结构基因将继续 使我们能够确定GLG的DNA序列分析（糖原合酶） 结构基因。 通过此分析，氨基酸序列将是 推论。 糖原生物合成生物合成的遗传调节 还将研究酶，希望GLG的隔离 大肠杆菌和鼠伤寒沙门氏菌的基因将为我们提供 在GLG表达表达的遗传调节模式下的一些见解 基因。