Deorphanizing the Peptidome

肽组去孤儿化

• 批准号: 8094525
• 负责人: IRIS LINDBERG
• 金额: $ 28.67万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-15 至 2013-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8094525
• 关键词: Biochemical; Bioinformatics; Brain; Cognition; Complement; Disease; Drug Delivery Systems; Enzymes; G Protein-Coupled Receptor Signaling; G-Protein-Coupled Receptors; Genome; Genomics; Health; Human Genome; In Vitro; Laboratories; Ligands; Mental Health; Mental disorders; Moods; Neurons; Neuropeptide Receptor; Neuropeptides; Neurotransmitters; Orphan; Pathway interactions; Peptide Receptor; Peptide Synthesis; Peptides; Physiological; Physiological Processes; Post-Translational Protein Processing; Prohormone Convertase; Proteins; Proteome; Reaction; Receptor Signaling; Research; Screening procedure; Signal Pathway; Signaling Molecule; Site; Technology; Testing; drug reward; human GPRC5C protein; innovation; novel; peptide G; receptor; secretory protein

The successful sequencing of the human genome has given rise to the next scientific opportunity of the twenty-first century: functional annotation of the proteome. About one-fifth of the genome encodes secretory proteins, a small number of which represent signaling molecules for G-protein coupled receptors (GPCRs). The major challenge that we will explore in this proposal is the comprehensive 'de-orphanization' (and thereby annotation) of the universe of peptides involved in neuronal GPCR signaling. There are currently about 100 non-olfactory orphan GPCRs, many of which are expected to use peptide ligands; however, fewer than a dozen novel peptides have been identified within the last eight years- and none at all within the last few years. Bioinformatics analyses indicate that the genome contains about 150 untested secretory proteins which possess biochemical similarities to known peptide precursors. We postulate that these proteins contain the missing peptide ligands for orphan GPCRs. However, in order to make these new orphan receptor- peptide matches, fresh approaches to peptide ligand identification are urgently needed. To identify novel peptide neurotransmitters we propose to take an innovative approach integrating expertise in bioactive peptide synthesis (Lindberg laboratory) with expertise in GPCR screening (Roth laboratory). We will experimentally confirm the presence of prohormone convertase-cleavable sites in a bioinformatically-derived list of putative precursors. Bioactive peptides will be generated from all validated precursors through large-scale in vitro posttranslational modification reactions using physiological enzymes (Lindberg laboratory). We will then discover cognate receptors to these peptides via functional screening against the entire genomic complement of known and orphan peptide receptors using facile screening technologies (Roth laboratory). Our results will enable us to match orphan receptors with novel peptide ligands, thus providing new neuropeptide-receptor signaling pairs. Since neuropeptide signaling pathways are critical to brain function and include pathways involved in mood and cognition, mental disorders, and drug reward, our results will significantly advance our understanding of mental health and disease, and may also generate new drug targets. While we will focus on obtaining and testing neuronally-expressed precursors/ligands and receptors, our research is also likely to uncover other ligand-receptor matches; thus a major impact on the many other physiological processes controlled by peptide- GPCR receptor signaling pathways is also anticipated.

人类基因组的成功测序引起了二十一世纪的下一个科学机会：蛋白质组的功能注释。大约五分之一的基因组编码分泌蛋白，其中少数代表了G蛋白偶联受体（GPCR）的信号分子。我们将在本提案中探索的主要挑战是涉及神经元GPCR信号涉及的肽宇宙的全面“脱键化”（从而注释）。目前大约有100个非野性孤儿GPCR，其中许多预计使用肽配体；但是，在过去的八年中，已经发现了不到十二种新颖的肽 - 在过去几年中根本没有发现。生物信息学分析表明，基因组包含约150种未经测试的分泌蛋白，这些蛋白质与已知肽前体具有生化相似性。我们假设这些蛋白质包含孤儿GPCR的缺失的肽配体。但是，为了使这些新的孤儿受体肽匹配，迫切需要新鲜的肽配体鉴定方法。为了确定新型肽神经递质，我们建议采用一种创新的方法，将生物活性肽合成（Lindberg Laboratory）与GPCR筛查（ROTH实验室）方面的专业知识相结合。我们将通过实验确认在生物信息衍生的假定前体列表中存在激素转化酶可转化位点的存在。生物活性肽将通过使用生理酶（Lindberg Laboratory）大规模的体外翻译后修饰反应从所有经过验证的前体产生。然后，我们将通过使用简便的筛选技术（ROTH Laboratory）对针对已知和孤儿肽受体的整个基因组补体进行功能筛选来发现这些肽的同源受体。我们的结果将使我们能够将孤儿受体与新型肽配体匹配，从而提供新的神经肽受体信号传导对。由于神经肽信号通路对大脑功能至关重要，包括情绪和认知，精神障碍和药物奖励涉及的途径，我们的结果将大大提高我们对心理健康和疾病的理解，还可能产生新的药物靶标。尽管我们将专注于获得和测试神经元表达的前体/配体和受体，但我们的研究也可能会发现其他配体受体匹配。因此，还预计对肽GPCR受体信号通路控制的许多其他生理过程的重大影响也被预计。