Structural Relay Methods for Functional Peptide Discovery

功能性肽发现的结构中继方法

基本信息

批准号：
8517720
负责人：
M.G. Finn
金额：
$ 20.11万
依托单位：
GEORGIA INSTITUTE OF TECHNOLOGY
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-08-01 至 2015-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8517720
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

项目摘要

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.

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，使蛋白质能够运输到细胞内、细胞之间、组织内和整个生物体中的适当位置。这种运输功能对于新的治疗和诊断药物的开发变得越来越重要，因为控制脱靶效应的要求变得越来越严格，并且递送的药物也越来越昂贵。然而，贩运很难做到，主要是因为它是一个多变量问题，因此超出了我们理性设计的能力。我们建议利用分子进化的强大工具来传递不携带遗传信息的试剂。我们将开发一种新的三步方法来识别将所需货物携带到细胞或特定细胞器中的肽。该方法依赖于 SELEX 技术（通过指数富集进行配体的系统进化）创建选择性结合任何特定肽的 DNA 序列的能力，以及噬菌体展示识别与任何 DNA 分子结合的肽的能力。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。该分子群中的一小部分预计会运输到所需的目的地，例如细胞质、线粒体或细胞核。该细胞材料的分离实现了所需肽与其余候选肽的分离，但分离量非常小。如果货物不编码肽的身份，目前还没有技术可以允许人们识别此类肽。我们希望SELEX和噬菌体展示技术的连续使用能够检测到功能肽并“放大”它们的身份信息，以便可以对它们进行识别、重新合成和测试。如果成功，这些研究应该会产生一种在各种环境和应用中发现功能性寡肽的通用方法。

项目成果

期刊论文数量（2）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（4）

Trimerization of the HIV Transmembrane Domain in Lipid Bilayers Modulates Broadly Neutralizing Antibody Binding.

DOI：
10.1002/anie.201508421
发表时间：
2016-02-18
期刊：
Angewandte Chemie (International ed. in English)
影响因子：
0
作者：
Reichart TM;Baksh MM;Rhee JK;Fiedler JD;Sligar SG;Finn MG;Zwick MB;Dawson PE
通讯作者：
Dawson PE

Polyvalent Catalysts Operating on Polyvalent Substrates: A Model for Surface-Controlled Reactivity.

在多价底物上运行的多价催化剂：表面控制反应性的模型。