ROLE OF INORGANIC POLYPHOSPHATES IN METABOLISM

无机多磷酸盐在新陈代谢中的作用

• 批准号: 3277238
• 负责人: HARLAND G. WOOD
• 金额: $ 22.19万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1981
• 资助国家: 美国
• 起止时间: 1981-07-01 至 1994-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3277238
• 关键词: Bacillus stearothermophilus; Propionibacteriaceae; biochemical evolution; chemical chain length; electron microscopy; enzyme mechanism; enzyme structure; gel electrophoresis; glucokinase; goats; laboratory rabbit; laboratory rat; liver cells; liver metabolism; liver regeneration; microorganism metabolism; phosphorylase kinase; polyphosphates; protein sequence; radiotracer; spectrometry; tissue /cell culture; yeasts

The immediate objective is to characterize the enzymes of polyphosphates (poly (P)) metabolism and then apply this information to the ultimate objective of elucidating the role of poly (P) in vivo. The studies will consist of 6 parts. (1) Polyphosphate kinase. A procedure has been developed by which poly (P) is clearly visible by electron microscopy. The aim is to investigate the binding of poly (P) is clearly visible by electron microscopy. The aim is to investigate the binding of poly (P) is clearly visible by electron microscopy. The aim is to investigate the binding of poly (P) to the enzyme and visualize the poly (P) and enzyme during the progressive lengthening of the chain. We also will investigate the amino acid sequence of the ATP binding site of poly (P) kinase. (2) Polyphosphate glucokinase. Propionibacterium shermanii and Nocardia minima are classified in the order Acetinomycetales which are considered to be primitive forms of life. Poly (P) glucokinase utilizes either poly (P) or ATP and N. minima contains another enzyme utilizing only ATP. N. minima may represent an example in a single organism of a transition from poly (P) to ATP as a phosphorylating agent. Glucokinase of Bacillus stearothermophilus, yeast hexokinase and of rat liver will also be investigated. The amino acid sequences will be determined using 5-p-fluorosulfonylbenzoyl adenosine as the labeling agent. The purpose will be to determine if there are similarities in the ATP sites which indicate an evolutionary continuity from ancient to more recent evolutionary forms. In addition, an attempt will be made to label and sequence the poly (P) site of poly (P) glucokinase. (3) Endopolyphosphatases. This enzyme or enzymes will be purified from yeast and it will be determined whether or not there is specificity for certain sizes of poly (P) and whether the cleavages yield certain lengths of poly (P). (4) Exopolyphosphatases. It has been shown that some factor is present in short-chain poly (P) which is required for the utilization of long-chain poly (P). This factor will be identified. (5) Role of poly (P) in vivo in P. shermanii. Using 32 P i , the poly (P) will be prelabeled by growing the bacteria in a lactate medium. The cells will be transferred a glucose medium containing unlabeled P i. The specific radioactivity of the glucose-6 poly (P) and the lambda-P of ATP will be determined to ascertain whether poly (P) serves as the phosphorylating agent in vivo. (6) Poly (P) in animal cells. Investigations will be continued to isolate and conclusively demonstrate that poly (P) is present in animals.

直接目标是表征多磷酸盐的酶 （poly（py（p））代谢，然后将此信息应用于最终 阐明聚（P）在体内作用的目的。 研究会 由6个部分组成。 （1）聚磷酸激酶。 一个程序已经 通过电子显微镜清楚地看到聚（P）的开发。 目的是研究poly（p）的结合。 电子显微镜。 目的是研究聚（P）的结合 通过电子显微镜可以清楚地看到。 目的是调查 聚（P）与酶的结合，并可视化聚（P）和 链条逐渐延长的酶。 我们也会 研究聚（P）的ATP结合位点的氨基酸序列 激酶。 （2）聚磷酸葡萄糖酶。 Shermanii丙泊菌和 Nocardia minima分为 被认为是生命的原始形式。 聚（P）葡萄糖激酶使用 poly（p）或ATP和N. minima含有另一种利用的酶 仅ATP。 N. minima可能代表了一个单个生物体的例子 从聚（P）到ATP的过渡为磷酸化剂。 葡萄糖激酶 芽孢杆菌嗜热，酵母己糖酶和大鼠肝脏 也可以调查。 氨基酸序列将使用 5-P-氟磺诺替氏腺苷作为标签剂。 目的 将确定AT​​P站点中是否有相似之处 表明从古代到最近的进化连续性 进化形式。 此外，将尝试标记和 序列聚（P）葡萄糖激酶的聚（P）位点。 （3） 肾磷酸酶。 该酶或酶将从 酵母将确定是否有特殊性 某些尺寸的聚（P）以及分裂是否产生一定的 聚（P）的长度。 （4）外聚磷酸酶。 已经表明 短链聚（P）中存在一些因素，这是 长链聚（P）的利用。 将确定此因素。 （5）聚（P）在体内的作用在shermanii中。 使用32 P I，Poly （P）将通过在乳酸培养基中种植细菌预先标签。 这 细胞将转移一个含有未标记P I的葡萄糖培养基。 葡萄糖-6聚（P）和lambda-p的特定放射性 将确定AT​​P确定Poly（P）是否用作 体内磷酸化剂。 （6）动物细胞中的聚（P）。 调查将继续隔离并最终证明 该聚（P）存在于动物中。