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PROLINE METABOLISM/REGULATION OF PURINES/ENERGY TRANSFER

脯氨酸代谢/嘌呤调节/能量转移

基本信息

批准号：
3295467
负责人：
DANIEL H KOHL
金额：
$ 14.71万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1987
资助国家：
美国
起止时间：
1987-07-01 至 1990-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/3295467
关键词：
Bacteroides Leguminoseae NAD(H) phosphate adenosine triphosphate amine oxidase (flavin) aminoacid metabolism biological signal transduction cell components cytoplasm electron transport nitrogen fixation nucleotide metabolism oxidation reduction reaction pentose phosphate shunt proline purine nucleotides spectrometry

项目摘要

We propose to examine the possible role of the proline metabolizing system in regulating energy-generating and biosynthetic pathways and in mediating intercompartmental energy transfer. Based on studies with erythrocytes and hepatocytes, it appears that pyrroline-5-carboxylate reductase (PCR) activity may modulate pentose phosphate pathway activity and result in the stimulation of de novo purine nucleotide biosynthesis. In addition PCR and proline oxidase (ProOx) may be involved in transfer of cytoplasmic redox potential into energy- utilizing organelles. Because of the uncommonly high levels of PCR and purine biosynthetic activity in nodules, the presence of ProOx in bacteroids (energy-consuming compartments lacking glycolytic capability), and the extremely high flux of energy transfer from the cytoplasm into bacteroids, nodules of Glycine max are an excellent model system to investigate the proposed roles of PCR and ProOx in intercompartmental energy transfer and purine nucleotide biosynthesis. The experimental plan is designed to examine the two different, but not mutually exclusive, functions of PCR and ProOx (i.e. regulation of purine nucleotide biosynthesis and energy transfer between organelles) proposed above. The specific objectives are: 1) to localize and characterize activities of PCR, ProOx, and other enzymes of proline metabolism; 2) to determine the relationship between PCR activity and purine nucleotide synthesis; 3) to define the role of proline in intercompartmental transfer of redox potential; and 4) to demonstrate the physiological significance of PCR and ProOx in intact tissues. The experimental approach involves biochemical characterization of PCR and ProOx, subcellular fractionation and tracer studies. The results obtained will contribute to our understanding of general mechanisms of cellular and subcellular energy transfer and metabolic regulation in eucaryotes.

我们建议研究脯氨酸的可能作用代谢系统调节能量产生和生物合成途径和介导房室间能量转移。基于红细胞和肝细胞，似乎吡咯啉-5-羧酸还原酶 (PCR) 活性可能调节磷酸戊糖途径活性并导致从头嘌呤核苷酸的刺激生物合成。此外，PCR 和脯氨酸氧化酶 (ProOx) 可能是参与细胞质氧化还原电位转化为能量利用细胞器。由于异常高的水平 PCR 和结节中嘌呤生物合成活性，存在类菌中的 ProOx（缺乏能量消耗区室）糖酵解能力），以及极高的能量通量从细胞质转移到类菌，大豆结节 max 是一个优秀的模型系统来研究所提出的 PCR 和 ProOx 在室间能量转移中的作用和嘌呤核苷酸生物合成。实验计划是旨在检查两者不同但不相互 PCR 和 ProOx 的独特功能（即嘌呤调节）核苷酸生物合成和细胞器之间的能量转移）上面提出的。具体目标是：1）本地化和表征 PCR、ProOx 和其他酶的活性脯氨酸代谢； 2）确定之间的关系 PCR活性和嘌呤核苷酸合成； 3）定义角色脯氨酸在氧化还原电位的间室转移中的作用；和 4）证明PCR的生理意义和完整组织中的 ProOx。实验方法包括 PCR 和 ProOx 的生化特征，亚细胞分馏和示踪剂研究。获得的结果将有助于我们理解细胞的一般机制以及亚细胞能量转移和代谢调节真核生物。