COMPLEMENT REGULATION BY APOLIPOPROTEIN J

长期目标是制定不同的调节方法， 末端补体复合物（TCC），以抑制其细胞溶解性 活性而不干扰需要亚裂解的活性 将复合物插入膜中，例如，在信号传导中。 这将有助于动脉粥样硬化和老年痴呆症等领域的研究 这两种疾病都涉及补体介导的细胞溶解。具体目标 1是确定载脂蛋白J（apoJ）- TCC相互作用， 对于体外TCC活性的缔合和抑制是重要的。到 为了解决这个问题，将进行实验以确定： apoJ和/或它的两个亚基中的任何一个是否干扰了 将任何单个组件组装到终端复合体中，以及2） 确定apoJ和/或其两个亚基中的任一个是否可以直接 抑制C9聚合。 具体目标2是鉴定apoJ序列 负责其与TCC的相互作用和抑制。 实验的目的是确定功能域，特别是 那些在α亚基，使用肽，并引入点 通过定点突变进入这些结构域，以鉴定 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.