NADPH-DEPENDENT METABOLISM OF ARACHIDONIC ACID

花生四烯酸的氧化代谢的特点是 其代谢途径和产物的复杂性不断增加， 归因于它们的功能特性的多样性。而新陈代谢 花生四烯酸分子模板的多功能性允许储存 以及转移一种特殊程度的有功能意义的化学物质 信息，这些多个反应的整合及其 与细胞、组织或身体相关的产物和生物活性 生理学是一项重要的任务，虽然困难和复杂，但 极大地促进了对生物化学的了解 反应，代谢物的结构和分子性质 所涉及的酶。微粒体细胞色素P-450花生四烯酸 环氧合酶催化四个区域异构体的对映选择性形成 环氧二十碳三烯酸(EETs)。EETs的非对称性 在几个组织中，内源性地展示了它们的酶 细胞色素P450为体内花生四烯酸的来源及确认 环氧合酶和，作为脂肪酸代谢级联的正式成员。 在过去的几年里，人们对描绘 细胞色素P-450在花生四烯酸生物活化中的作用 因此，在其对于细胞和器官生理学的意义上。许多. 这些努力受到了强效生物活性的刺激 EETs作为多肽激素释放、血管活性分子的介体 以及膜离子通量的调节剂。最近，有几行 有证据表明，这条途径可能以一种尚未确定的方式参与其中， 在高血压的病理生理学方面。潜在的生理和 这一建议的临床应用具有关键的重要性和范围。 因此，激发了广泛的研究兴趣。 在过去的10年里，这个项目完成了详细的 该酶的酶学性质和结构性质的研究 环氧合酶代谢物，这些研究对我们的 目前对其生物化学和功能意义的认识 这条路。进一步的进展需要详细的分子知识。 所涉及的酶，以及生物特异性的发展 在蛋白质水平研究这一途径的调控所需的探针 和基因水平。我们建议利用现代分子生物学技术 为了鉴定和鉴定细胞色素P-450亚型 参与内源性EET的产生。我们建议克隆 相关基因(S)，表达所克隆的cDNAs，以鉴定 重组蛋白(S)，制备免疫特异性探针，并研究 重组蛋白在细胞内皮细胞和钙代谢中的作用

The oxygenated metabolism of arachidonic acid is characterized by the increasing complexity of its metabolic pathways and products and, by the diversity of functional properties attributed to them. While the metabolic versatility of the arachidonate molecular template allows for the storage and transfer of an exceptional degree of functionally meaningful chemical information, the integration of these multiple reactions and of their associated products and biological activities to cell, tissue or body physiology is an important task which, although difficult and complex, is greatly facilitated by the understanding of the biochemistry of the reactions, the structure of metabolites and the molecular properties of the enzymes involved. The microsomal cytochrome P-450 arachidonic acid epoxygenase catalyzes the enantioselective formation of four regioisomeric epoxyeicosatrienoic acids (EETs). The asymmetric nature of the EETs present, endogenously, in several tissues demonstrated their enzymatic origin and confirmed cytochrome P450 as the in vivo arachidonic acid epoxygenase and, as a formal member of the fatty acid metabolic cascade. The last few years have seen a renewed interest in the delineation of the role of cytochrome P-450 in the bioactivation of arachidonic acid and, therefore, in its significance for cell and organ physiology. Many of these efforts have been stimulated by the potent biological activities of the EETs as mediators for peptide hormone release, vasoactive molecules and modulators of membrane ion fluxes. More recently, several lines of evidence suggest that this pathway may be involved, in yet undefined ways, in the pathophysiology of hypertension. The potential physiological and clinical implication of this proposal are of pivotal importance and area therefore, stimulating extensive research. During the last 10 years, this project completed a detailed characterization of the enzymology and the structural properties of the epoxygenase metabolites, those studies contributed substantially to our present understanding of the biochemical and functional significance of this pathway. Further progress will require a detailed molecular knowledge of the enzymes involved, as well as the development of the biospecific probes needed for studies of the regulation of this pathway at the protein and gene level. We propose to utilize modern molecular biology techniques in order to identify and characterize the cytochrome P-450 isoforms involved in the generation of endogenous EETs. We propose to clone the relevant gene(s), to express the cloned cDNA to characterize the recombinant protein(s), to generate immunospecific probes and to study the role of recombinant proteins in cell EET and calcium metabolism.