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，使蛋白质能够传播到ELL，ELL，细胞之间，组织中以及整个生物体中的适当位置。这种贩运功能对于新的治疗和诊断剂的开发越来越重要，因为控制脱靶效应的要求变得越来越严格，并且交付的代理商越来越昂贵。 但是，贩运很难做，主要是因为这是一个多元问题，因此超出了我们的理性设计能力。我们建议携带强大的分子进化工具，以承担不携带遗传信息的药物的传递。我们将开发一种新的三步方法，以鉴定将所需货物携带到细胞或特定细胞细胞器中的肽。该方法依赖于SELEX技术（通过指数富集对配体的系统演化）创建与任何特定肽结合的DNA序列的能力，以及噬菌体显示出鉴定与任何DNA分子结合的肽的能力。 使用此类工具，我们将准备与感兴趣的货物绑定的肽库，并将其与感兴趣的细胞孵化。可以预期，该分子种群的很小一部分可以传播到所需的目的地，例如细胞质，线粒体或核。该细胞材料的分离实现了所需肽与其余候选物的分离，但量很少。目前没有任何技术可以允许人们识别此类肽，如果货物没有为肽的身份进行编码。我们希望SELEX和噬菌体显示技术的顺序使用能够检测到功能性肽，并“放大”其身份的信息，以便可以识别，重新合成和测试。 如果成功，这些研究应导致一种在各种环境和应用中发现功能性寡肽的一般方法。 公共卫生相关性：现代药物开发需要可以将治疗剂传递给体内特定细胞的分子。我们建议使用化学合成和分子进化方法的组合来测试一种发现具有这种能力的肽分子的新方法。最终的技术应允许几乎所有药物靶向任何所需的生物学目的地。