Identification of Novel Peptide Hormones

描述（申请人提供）：分泌蛋白基因组编码的五分之一；这些蛋白质中的一小部分代表肽激素信号分子。这一建议涉及识别新的分泌分子参与代谢功能的控制。由于大多数神经内分泌信号分子经历蛋白转化酶介导的成熟，我们计划利用新前体中真正可切割位点的存在来验证一组潜在的肽前体。然后，我们将测试从这些蛋白质中获得的正确修饰肽产品的生物活性。这些检测将识别生物活性肽作用于组织是代谢控制的关键效应器：脂肪，肝脏和肌肉。拟议的项目代表了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对大量假定的前体进行生物信息学鉴定，然后用纯化的重组转化酶筛选hek表达蛋白（Lindberg实验室）。然后，由加工筛选产生的信息将用于指导林德伯格实验室中有希望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公司进行针对脂肪、肝脏和肌肉细胞葡萄糖代谢的六种代谢试验。我们期望这些研究将导致鉴定出几种有助于代谢控制的新型肽激素。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，重点是鉴定新的生物信息学鉴定的肽激素。我们将使用生理酶对候选前体进行化学处理，制成肽混合物，我们将在六种不同的葡萄糖和脂质代谢试验中进行测试。我们期望这些研究将导致鉴定出几种有助于代谢控制的新型肽激素。

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.