COMPUTATIONAL METHODS FOR ANALYSIS OF INTERMEDIARY METABOLISM

中间代谢分析的计算方法

• 批准号: 6205880
• 负责人: F MARK JEFFREY
• 金额: $ 6.95万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-08-15 至 2000-08-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6205880
• 关键词: Mammalia; animal tissue; biomedical resource; cardiovascular system; computers; human tissue; liver; nuclear magnetic resonance spectroscopy; statistics /biometry; technology /technique

This project involves the development of mathematical models to estimate the flux through different pathways using isotopomer analysis and 13C NMR data. Three programs are under development: tcaSIM, a simulation of the citric acid cycle and related pathways which generates 13C isotopomer data for use in the design of experiments; tcaCALC, a model which estimates relative pathway fluxes from NMR spectra obtained at metabolic and isotopic steady state; and tcaFLUX, a kinetic analysis of the citric acid cycle which allows measurement of absolute flux from systems at metabolic, but not isotopic, steady state. There has been much progress in the development of the kinetic analysis. It has been applied to multiplet data and fractional enrichment data collected from intact hearts and after freeze-clamping. A number of different substrate groups were examined. The model can be used to estimate citric acid cycle activity, the exchange between citric acid cycle intermediates and amino acids (which will include transportation between different cellular compartments), the contribution of labeled substrate to acetyl-CoA, and anaplerosis. The citric acid cycle rate determined by the model was used to estimate oxygen consumption; the values obtained were similar to those measured directly. It has been found that the use of multiplet data along with standard fractional carbon enrichments in this analysis is beneficial, reducing the correlation between parameters that otherwise exists. There has also been success with the application of data available from GCMS to isotopomer analysis. A Finnegan GCQ bench-top GCMS with tandem mass spectrometry capability is in use. This allows the investigator to acquire, in addition to standard full-scan mass spectra, mass spectra of parent ions which have been fragmented after inducing collisions with the carrier gas. The mass spectra of these fragments give additional information on the isotopomers present in the metabolite mixture. Using the methyl-8 derivative of glutamate isolated from hearts perfused under a variety of conditions, such data was analyzed in a similar fashion to NMR multiplet data to determine substrate utilization and anaplerosis. The results were very similar to those obtained using NMR data, indicating that this technique will be useful as a more-sensitive, complementary approach to NMR. (Core 3) REPORT PERIOD: (09/01/97-08/31/98)

该项目涉及开发数学模型 使用同位素分析估算通过不同途径的通量 和13C NMR数据。 正在开发三个程序：tcasim，一个 柠檬酸周期和相关途径的模拟 生成13C同位素数据以用于实验设计； TCACALC，该模型估算NMR的相对途径通量 在代谢和同位素稳态下获得的光谱；和TCAFLUX， 柠檬酸周期的动力学分析，该循环允许测量 在代谢中的系统中的绝对通量，但不是同位素，稳定的 状态。 动力学的发展取得了很大进展 分析。 它已应用于多重数据和分数 从完整心脏和之后收集的富集数据 冻结夹。 许多不同的基板组是 检查。 该模型可用于估计柠檬酸周期 活动，柠檬酸循环中间体之间的交换 氨基酸（其中包括不同的运输 细胞室），标记的底物对 乙酰辅酶A和腐败。 由 该模型用于估计消耗氧。获得的值 与直接测量的相似。 已经发现 使用多重数据以及标准分数碳 该分析的富集是有益的，可以减少相关性 在否则存在的参数之间。 也有成功 随着GCM的可用数据的应用到同位素 分析。 Finnegan GCQ台式GCM具有串联质谱法 功能正在使用。 这使调查员可以在 除标准全扫描质谱，父级的质谱 诱导与 载气。 这些片段的质谱给出了额外的 有关代谢物混合物中存在的同位素剂的信息。 使用从心脏分离的谷氨酸的甲基-8衍生物 在各种条件下灌注，在A中分析了此类数据 与NMR多重数据类似的方式以确定基板 利用和孤立性。 结果与那些非常相似 使用NMR数据获得，表明此技术将很有用 作为一种更敏感的NMR互补方法。 （核心3）报告 周期：（09/01/97-08/31/98）