COMPLEMENT REGULATION BY APOLIPOPROTEIN J

长期目标是开发不同的方法来监管 末端补体复合体(TCC)以抑制其细胞溶解 活动而不干扰需要亚分辨率的活动 将该复合体插入膜中，例如在信号转导中。 这将对动脉粥样硬化和阿尔茨海默氏症等领域有所帮助 疾病，两者都涉及补体介导的细胞溶解。特定目标 L将确定载脂蛋白J(ApoJ)与TCC的相互作用 对体外联合和抑制TCC活性具有重要意义。至 针对这个问题，将进行实验，以确定L) 载脂蛋白J和/或其两个亚基中的任何一个是否干扰 将任何单个组件组装到终端综合体中，以及2) 确定apoJ和/或其两个亚基中的任何一个是否可以直接 抑制C9聚合。具体目标2是鉴定apoJ序列 对其与TCC的相互作用和对其的抑制负责。 实验旨在确定功能域，特别是 那些在阿尔法亚基中，使用多肽，并介绍点 通过定点突变识别这些结构域的突变 功能性氨基酸残基。

The long term objective is to develop ways to differentially regulate the terminal complement complex (TCC) in order to inhibit its cytolytic activity without interfering with activities that require sublytic insertion of the complex into membranes, e.g., in signal transduction. This would be helpful in such areas as atherosclerosis and Alzheimer's disease, both of which involve complement-mediated cytolysis. Specific aim l is to determine the apolipoprotein J (apoJ) - TCC interactions that are important for the association and inhibition of TCC activity in vitro. To address this question, experiments will be performed to l) ascertain whether apoJ and/or either of its two subunits interferes with the assembly of any individual component into the terminal complex, and 2) determine whether apoJ and/or either of its two subunits can directly inhibit C9 polymerization. Specific aim 2 is to identify apoJ sequences responsible for its interaction with and inhibition of the TCC. Experiments are designed to identify functional domains, specifically those in the alpha subunit, using peptides, and to introduce point mutations into these domains via site-directed mutagenesis to identify functional amino acid residues.