Structural Relay Methods for Functional Peptide Discovery

功能性肽发现的结构中继方法

• 批准号: 8359132
• 负责人: M.G. Finn
• 金额: $ 15.15万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-08-01 至 2013-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8359132
• 关键词: Adopted; Binding; Biological; Biological Products; Biology; Biotin; Cell Nucleus; Cells; Code; Collecting Cell; Cytoplasm; DNA; DNA Sequence; Destinations; Development; Dreams; Evolution; Genetic; Hand; Human Cell Line; Incubated; Inferior; Laboratories; Laboratory Scientists; Lead; Libraries; Ligands; Location; Methods; Mind; Mitochondria; Molecular; Molecular Evolution; Oligopeptides; Organelles; Organism; Peptide Library; Peptide Phage Display Library; Peptides; Phage Display; Pharmaceutical Preparations; Polynucleotides; Population; Property; Proteins; Research Personnel; Rest; Ribosomes; Science; Side; Structure; Techniques; Technology; Testing; Therapeutic Agents; Tissues; Ursidae Family; aptamer; base; chemical synthesis; combinatorial chemistry; design; drug development; interest; novel diagnostics; novel therapeutics; operation; protein aminoacid sequence; small molecule; success; tool; trafficking; uptake

DESCRIPTION (provided by applicant): Oligopeptides have many functions in biology, including providing binding selectivity that allows proteins to traffic to the proper place in the ell, between cells, within tissues, and throughout the organism. This trafficking function is becoming more and more important to the development of new therapeutic and diagnostic agents, as the requirements for the control of off-target effects are becoming ever more stringent and the delivered agents ever more expensive. However, trafficking is hard to do, mostly because it is a multivariate problem, and so is beyond our capabilities of rational design. We propose to bring powerful tools of molecular evolution to bear on the delivery of agents that do not carry genetic information. We will develop a new three-step method to identify peptides that carry desired cargoes into cells or to particular cellular organelles. The method relies on the ability of the SELEX technique (systematic evolution of ligands by exponential enrichment) to create DNA sequences that bind selectively to any particular peptide, and the ability of phage display to identify peptides that bind to any DNA molecule. With such tools in hand, we will prepare libraries of peptides bound to the cargo of interest, and incubate them with cells of interest. A very small fraction of that molecular population can be expected to traffic to the desired destination, such as the cytoplasm, mitochondria, or nucleus. Isolation of that cellular material accomplishes the separation of the desired peptide(s) from the rest of the candidates, but in very small amounts. No technique is currently available that will allow one to identify such peptides if the cargo does not code for the identity of the peptide. We hope that the sequential use of the SELEX and phage display techniques will allow the functional peptides to be detected and the information of their identities "amplified" so that they can be identified, resynthesized, and tested. If successful, these studies should lead to a general method to discover functional oligopeptides in a wide variety of settings and applications. PUBLIC HEALTH RELEVANCE: Modern drug development requires molecules that can deliver therapeutic agents to specific cells in the body. We propose to test a new method for discovering peptide molecules that have this ability, using a combination of chemical synthesis and methods of molecular evolution. The resulting technique should allow almost any drug to be targeted to any desired biological destination.

描述（由申请人提供）：寡肽在生物学中具有许多功能，包括提供结合选择性，允许蛋白质运输到细胞内、细胞间、组织内和整个生物体中的适当位置。这种运输功能对于开发新的治疗和诊断剂变得越来越重要，因为对控制脱靶效应的要求变得越来越严格，并且递送的药剂越来越昂贵。 然而，贩卖人口很难做到，主要是因为它是一个多变量的问题，因此超出了我们理性设计的能力。我们建议使用强大的分子进化工具来传递不携带遗传信息的药物。我们将开发一种新的三步法来鉴定携带所需货物进入细胞或特定细胞器的肽。该方法依赖于SELEX技术（通过指数富集的配体系统进化）产生选择性结合任何特定肽的DNA序列的能力，以及噬菌体展示鉴定结合任何DNA分子的肽的能力。 有了这些工具，我们将准备与感兴趣的货物结合的肽库，并将它们与感兴趣的细胞一起孵育。可以预期该分子群体的非常小的部分运输到期望的目的地，例如细胞质、线粒体或细胞核。细胞物质的分离完成了所需肽与其余候选物的分离，但量非常小。如果货物不编码肽的身份，目前没有技术可以允许鉴定这样的肽。我们希望，SELEX和噬菌体展示技术的顺序使用将允许功能肽被检测和它们的身份的信息“放大”，以便它们可以被识别，重新合成和测试。 如果成功的话，这些研究应该会导致一个通用的方法，以发现功能性寡肽在各种各样的设置和应用。 公共卫生相关性：现代药物开发需要能够将治疗剂递送到体内特定细胞的分子。我们建议测试一种新的方法来发现具有这种能力的肽分子，使用化学合成和分子进化方法的组合。由此产生的技术应该允许几乎任何药物靶向任何所需的生物目的地。