Identification of Novel Peptide Hormones

描述（由申请人提供）：基因组的五分之一编码分泌蛋白;这些蛋白质的一小部分代表肽激素信号分子。这一建议解决了识别新的分泌分子参与控制代谢功能。由于大多数神经内分泌信号分子经历前蛋白转化酶介导的成熟，我们计划利用新的前体中存在的真正可裂解的位点来验证一组潜在的肽前体。然后，我们将测试从这些蛋白质衍生的正确修饰的肽产物的生物活性。这些检测将鉴定作用于作为代谢控制的关键效应物的组织的生物活性肽：脂肪、肝脏和肌肉。该项目代表了Lindberg实验室与分泌蛋白公司Five Prime Therapeutics的合作。简而言之，通过Five Prime对大量推定前体进行生物信息学鉴定，然后用纯化的重组转化酶（Lindberg实验室）筛选HEK表达的蛋白质。然后，处理屏幕产生的信息将用于指导Lindberg实验室中有前途的前体的大规模重组蛋白生产。这些纯化的前体将在Lindberg实验室进行体外蛋白水解和终末成熟反应，然后在FivePrime Therapeutics针对脂肪，肝脏和肌肉细胞葡萄糖代谢的六种代谢试验中测试所得肽产物。我们希望这些研究将导致几种新的肽激素有助于代谢控制的鉴定。这些结果应该有助于我们全面了解激素对葡萄糖和脂质代谢的控制。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肽激素。我们将使用生理酶对候选前体进行化学处理，以制备肽混合物，我们将在六种不同的葡萄糖和脂质代谢测定中进行测试。我们希望这些研究将导致几种新的肽激素有助于代谢控制的鉴定。

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.