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(二十烷基类)表达中的作用：这项建议的总体目标是确定特定的红藻次级天然产物二十烷基类参与这些生物的应激反应(通过应激蛋白的产生或与应激蛋白的产生相关)。二十烷类化合物是一种生物活性分子，参与人体许多基本的代谢功能，在哺乳动物体内平衡中发挥重要作用，它们几乎存在于哺乳动物的每一个组织和体液中，并发挥着广泛的生理活动。负责二十烷类新陈代谢的酶系统在哺乳动物中出现之前就进化了100‘S。确定二十烷类化合物进化为压力管理化合物，并且它们的作用通过进化得到保守，这将提供与其在动物中的主要作用之一的功能同源性，在动物中，脂氧合酶介导的白三烯和脂氧素的形成是启动有机体内应激反应的信号。这些目标将通过在实验室培养目标物种Periclados，并检测应激蛋白表达、脂氧合酶诱导和随后的二十烷类化合物表达的时间进程来实现。应激蛋白的鉴定将采用双向SDS-PAGE。二十烷类化合物的浓度将使用酶联免疫吸附试验进行测量。将开发一种针对莫雷氏菌脂肪氧合酶的分子探针，所产生的基因或基因片段将被用于生产多克隆抗体。这些抗体随后将用于应激条件下脂氧合酶的诱导，方法是采用ELISA法。

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.