ROLE OF STRESS IN EXPRESSION OF MAMMALIAN EICOSANOIDS IN RED ALGAE

应激在红藻中哺乳动物次级激素（类花生酸）表达中的作用：本提案的总体目标是确定特定的红藻次级天然产物，类花生酸参与这些生物体中的应激反应（通过应激蛋白的产生发出信号或与应激蛋白的产生相关）。类二十烷酸是参与许多基本人类代谢功能的生物活性分子，在哺乳动物体内平衡中发挥重要作用，其中它们存在于几乎每个哺乳动物组织和体液中，并且它们发挥广泛的生理活性。负责类花生酸代谢的酶系统在哺乳动物中出现之前进化了数百万年。类二十烷酸作为应激管理化合物进化，并且它们的作用通过进化而被保守，这一点的确立将提供与其在动物中的主要作用之一的功能同源性，其中脂氧合酶介导的白三烯和脂氧素的形成用作在生物体内引发应激反应的信号。目标将通过在实验室中培养目标物种Murrayella periclados并检查应激蛋白表达、脂氧合酶诱导和随后的类花生酸表达的时间过程来实现。强制降解蛋白鉴定将采用二维SDS PAGE。将使用ELISA测定法测量类花生酸浓度。将开发Murrayella脂氧合酶的分子探针，并将所得基因或基因片段用于生产多克隆抗体。然后将抗体用于在应激条件下使用ELISA测定法跟踪脂氧合酶诱导。

Role of Stress in Expression of Mammalian Secondary Hormones (Eicosanoids) in Red Algae: The overall objective of this proposal is to establish that specific red algal secondary natural products, eicosanoids are involved in stress response (signaled by or correlated with the production of stress proteins) in these organisms. Eicosanoids are biologically active molecules that are involved in many basic human metabolic functions, playing important roles in mammalian homeostasis where they are present in virtually every mammalian tissue and body fluid and where they exert a broad spectrum of physiological activities. Enzyme systems responsible for eicosanoid metabolism evolved 100's of millions of years prior to their appearance in mammals. Establishment that eicosanoids evolved as stress management compounds, and that their role has been conserved through evolution would provide a functional homology with one of its major roles in animals where the lipoxygenase- mediated formation of leukotrienes and lipoxins serves as a signal to initiate stress responses within the organisms. The objectives will be carried out by culturing the target species, Murrayella periclados, in the laboratory and examining the time course of stress protein expression, lipoxygenase enzyme induction and subsequent eicosanoid expression. Stress protein identification will employ 2 dimensional SDS PAGE. Eicosanoid concentrations will be measured using ELISA assays. A molecular probe for Murrayella lipoxygenase will be developed and the resultant gene or gene fragment will be utilized to produce polycolona antibodies. The antibodies will then be used to follow lipoxygenase induction following stress conditions utilizing ELISA assays.