Mechanism and physiology of DUF1222-assisted lipase maturation

产品描述： 脂蛋白脂酶（LPL）水解甘油三酯是维持正常血脂所必需的 新陈代谢.如果没有新鉴定的， 跨膜蛋白、脂肪酶成熟因子（LMF 1）。LMF 1基本上是 事实上，它属于一个蛋白质家族，具有类似的“结构域”， 未知函数（Duf 1222）。该项目的主要目标是调查 LPL和LMF 1之间的相互作用。通过类比具有结构特征的脂肪酶 与LPL类似，我们提出LMF 1作为LPL的“私人折叠酶”。在目标1中，我将 使用生化分析和细胞分析来确定LPL需要折叠的原因 LMF 1如何帮助LPL折叠。目的2包括膜的分析 LMF 1的拓扑和结构。最后，目标3侧重于观察到， 人类病原体有Duf 1222蛋白，因此我将使用计算和 生物化学方法来确定Duf 1222蛋白是否代表了一种 折叠酶-脂肪酶对。在指导阶段，我将在我的初步 观察，以开发LMF 1膜拓扑结构的模型。另外我将开始 LMF-LPL相互作用的生化和结构分析，并开始初步 努力实现目标3。在独立阶段，我将继续对LMF 1进行结构分析 并将我的生化分析扩展到其他物种中的Duf 1222蛋白，以确定 它们在脂质代谢中具有重要作用。

Description: Hydrolysis of triacylglycerols by lipoprotein lipase (LPL) is essential for normal lipid metabolism. LPL does not fold and function properly without a newly identified, transmembrane protein, lipase maturation factor (LMF1). LMF1 is essentially uncharacterized; in fact it belongs to a family of proteins with a similar "domain of unknown function" (Duf1222). The major goal of this project is to investigate the interaction between LPL and LMF1. By analogy to lipases with structural characteristics similar to LPL, we propose that LMF1 acts as a "private foldase" for LPL. In Aim 1, I will use biochemical assays and assays in cells to determine why LPL needs folding assistance and how LMF1 helps LPL fold. Aim 2 consists of analysis of the membrane topology and structure of LMF1. Finally, Aim 3 focuses on the observation that many human pathogens have Duf1222 proteins; therefore I will use computational and biochemical approaches to determine if the Duf1222 protein represents one-half of a foldase-lipase pair. During the mentored phase, I will build on my preliminary observations to develop a model for LMF1 membrane topology. Additionally I will begin biochemical and structural analyses of the LMF-LPL interaction, and begin preliminary work on aim 3. For the independent phase, I will continue structural analysis of LMF1 and expand my biochemical analysis to Duf1222 proteins in other species to determine if they have important roles in lipid metabolism.