Identification of Novel Peptide Hormones

描述(申请人提供)：五分之一的基因组编码分泌蛋白质；这些蛋白质中的一小部分代表多肽激素信号分子。这项建议致力于鉴定参与代谢功能控制的新的分泌分子。由于大多数神经内分泌信号分子都经历了原蛋白转换酶介导的成熟，我们计划利用新前体中真实可切割位点的存在来验证一系列潜在的肽前体。然后，我们将测试从这些蛋白质中提取的正确修饰的多肽产品的生物活性。这些分析将识别作用于代谢控制关键效应器的组织：脂肪、肝脏和肌肉的生物活性多肽。拟议的项目是林德伯格实验室与分泌性蛋白质公司Five Prime Treeutics的合作。简而言之，用五素对大量假定的前体进行生物信息学鉴定之后，将用纯化的重组转化酶筛选HEK表达的蛋白(林德伯格实验室)。然后，处理筛选产生的信息将被用于指导林德伯格实验室大规模生产有希望的前体的重组蛋白。这些纯化的前体将在林德伯格实验室进行体外蛋白分解和最终成熟反应，然后在FivePrime Treeutics针对脂肪、肝脏和肌肉细胞的葡萄糖代谢进行六项代谢测试。我们期望这些研究将有助于确定几种有助于代谢控制的新的多肽激素。这些结果应该有助于我们完全理解激素对糖和脂代谢的控制。公共卫生相关性：这是一个发现项目，重点是识别新的生物信息学识别的多肽荷尔蒙。我们将使用生理酶对候选前体进行化学处理，以制造多肽混合物，我们将在六种不同的葡萄糖和脂肪代谢分析中进行测试。我们期望这些研究将有助于确定几种有助于代谢控制的新的多肽激素。

DESCRIPTION (provided by applicant): One-fifth of the genome codes for secretory proteins; a small subset of these proteins represent peptide hormone signaling molecules. This proposal addresses the identification of novel secretory molecules involved in the control of metabolic function. Since the majority of neuroendocrine signaling molecules undergo proprotein convertase-mediated maturation, we plan to exploit the presence of genuinely cleavable sites in novel precursors to validate a set of potential peptide precursors. We will then test the bioactivity of correctly modified peptide products derived from these proteins. These assays will identify biologically active peptides acting on tissues that are key effectors of metabolic control: fat, liver and muscle. The proposed project represents a collaboration of the Lindberg laboratory with the secretory protein company Five Prime Therapeutics. Briefly, bioinformatics identification of large numbers of putative precursors by Five Prime will be followed by screening of HEK-expressed proteins with purified recombinant convertases (Lindberg laboratory). The information generated by the processing screen will then be used to direct large-scale recombinant protein production of promising precursors in the Lindberg laboratory. These purified precursors will be subjected to in vitro proteolytic and terminal maturation reactions in the Lindberg laboratory, and the resulting peptide products will then be tested in six metabolic assays targeted at fat, liver and muscle cell glucose metabolism at FivePrime Therapeutics. We expect that these studies will result in the identification of several novel peptide hormones contributing to metabolic control. These results should aid in our complete understanding of the hormonal control of glucose and lipid metabolism. PUBLIC HEALTH RELEVANCE: This is a discovery project which focuses on the identification of novel bioinformatics-identified peptide hormones. We will employ chemical processing of candidate precursors using physiological enzymes to make peptide mixtures which we will test in six different assays of glucose and lipid metabolism. We expect that these studies will result in the identification of several novel peptide hormones contributing to metabolic control.