NADPH-DEPENDENT METABOLISM OF ARACHIDONIC ACID

花生四烯酸的氧合代谢的特征在于： 其代谢途径和产物的复杂性增加， 多样性的功能属性归因于他们。虽然新陈代谢 花生四烯酸分子模板的多功能性允许储存 和转移功能上有意义的化学物质的特殊程度 信息，这些多重反应的整合， 相关产品和生物活性对细胞、组织或身体的影响 生理学是一项重要的任务，虽然困难和复杂， 对生物化学的理解， 反应，代谢产物的结构和分子特性， 所涉及的酶。微粒体细胞色素P-450花生四烯酸 环氧合酶催化四种区域异构体的对映选择性形成， 环氧二十碳三烯酸（ESTs）。埃克塞特的不对称性 目前，内源性地，在几个组织证明了他们的酶 来源并确认细胞色素P450为体内花生四烯酸 环氧合酶，并作为脂肪酸代谢级联的正式成员。 最近几年，人们对划定 细胞色素P-450在花生四烯酸生物活化中的作用， 因此，它对细胞和器官生理学的意义。许多 这些努力是由以下物质的有效生物活性所激发的： 作为肽类激素释放、血管活性分子 和膜离子通量的调节剂。最近，几行 有证据表明，这一途径可能以尚未确定的方式， 高血压的病理生理学。潜在的生理和 这一建议的临床意义是至关重要的， 因此，激发了广泛的研究。 在过去的10年里，该项目完成了详细的 的酶学和结构特性的表征， 环氧酶代谢物，这些研究大大有助于我们的研究。 目前对生物化学和功能意义的理解， 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 在蛋白质中研究该途径的调节所需的探针 基因水平。我们建议利用现代分子生物学技术 为了鉴定和表征细胞色素P-450亚型， 参与了内源性雌二醇的产生。我们建议克隆 相关基因，以表达克隆的cDNA来表征 重组蛋白，以产生免疫特异性探针并研究 重组蛋白在细胞EET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.