INTERACTION OF SARCOSINE OXIDASE W/ (METHYLSELENO)ACETATE

肌氨酸氧化酶与（甲基硒）乙酸的相互作用

• 批准号: 6120839
• 负责人: MARILYN S JORNS
• 金额: $ 3.95万
• 依托单位: UNIVERSITY OF CALIF-LOS ALAMOS NAT LAB
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-01-15 至 2000-01-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6120839
• 关键词: biological products; biomedical resource; proteins

The SIR provided (2S)-N-(2'-O-benzyl-glycoloyl)bomane-10-,2-sultam 11,2- 13C]acetic acid; 20g Recently, a new pathway to isopentenyl pyrophosphate has been discovered which emanates from carbohydrate metabolism. To probe distribution of this new pathway in nature and to elucidate further intermediates in the pathway, we are presently synthesizing 1-deoxy-D-[2,4-13C2]xylulose. Initially, a small quantity for exploratory work will be prepared from ethyl bromo- [ I - I 3C] acetate and [2-13C]acetone, which has been developed by the UW group. In the course of this work it became evident that this route is not very efficient and thus unsuitable for the production of larger quantities of this precursor. A second route has therefore been designed by the SIR, which makes effective use of the labeled synthons they have developed. If experiments on ubiquinone formation in E. coli with the initially synthesized precursor sample demonstrate the feasibility of this approach, a larger quantity of the compound will then be synthsized by the SIR. This precursor will be fed to a variety of biological systems synthesizing isoprenoid compounds and the products will be analyzed by 13C-NMR for the incorporation of 13C and the appearance of 13C_13C coupling, resulting from the rearrangement of the precursor, in the appropriate positions. This will give an overview of the relative importance of the new pathway compared to the classical mevalonate pathway. Next, we will incubate the double labeled substrate with washed cells and cell-free extracts of E. coli directly in the NMR spectrometer and examine the incubations periodically by 1H-detected 13C-NMR, looking specifically for double-quantum transitions, for the appearance of new species in which the two 13C nuclei are connected to each other. Since virtually no other metabolic pathway can give rise to extensive 13C-13C coupling, this will be a very specific probe for the intermediates in the new pathway of isoprenoid biosynthesis. The time course of the appearance and disappearance of new species will indicate their relative position in the reaction sequence, and labeling of the precursor in additional positions will allow the assignment of structures to these intermediates. These in vivo experiments will require high sensitivity to detect multiquantum transitions from very small amounts of transient compounds. This can be achieved on the 750 MHz NMR spectrometer at the Univ. of Wash.

SIR提供的（2s）-n-（2'-O-苯二唑 - 糖果）Bomane-10-，2-Sultam 11,2-13C]乙酸； 20G最近，一条新的等烯基途径 已经发现了焦磷酸盐从碳水化合物中散发出来的 代谢。 探测自然界的这种新途径的分布 为了阐明途径中进一步的中间体，我们目前是 合成的1-脱氧-D- [2,4-13C2]木糖。 最初，一个小 探索性工作的数量将由乙基bromo- [i- I 3C]乙酸盐和[2-13c]丙酮，已由UW开发 团体。 在这项工作的过程中，很明显这条路线 不是很有效，因此不适合生产较大 该前体的数量。 因此，第二条路线已经 由SIR设计，有效地使用标记的合成子 他们已经发展了。 如果在E中进行泛氨基酮形成的实验。 最初合成的前体样品的大肠杆菌证明了 这种方法的可行性，大量的化合物将 然后由先生合成。 这个前体将被喂给 各种生物系统合成的类异戊二烯化合物和 该产品将通过13C-NMR分析13C的掺入 以及13C_13C耦合的外观 前体的重排，处于适当的位置。 这 将概述新途径的相对重要性 与经典的甲谷酸途径相比。 接下来，我们将孵化 带有洗涤细胞和无细胞提取物的双重标记底物 直接在NMR光谱仪中的大肠杆菌 定期通过1H检测的13C-NMR孵育，专门查看 对于双重量子过渡，用于出现新物种 两个13C核相互连接。 从虚拟上 没有其他代谢途径可以引起广泛的13C-13C 耦合，这将是对中间体的非常特殊的探针 类异丙生素生物合成的新途径。 时间课程的时间课程 新物种的外观和消失将表明他们 反应序列中的相对位置，并标记 前体处于其他职位，将允许分配 这些中间体的结构。 这些体内实验将 需要高灵敏度才能检测到非常敏感的过渡 少量的瞬态化合物。 这可以在750上实现 大学的MHz NMR光谱仪。 清洗。