Genetically Encoded Smart Biohybrid Materials

基因编码智能生物混合材料

• 批准号: 10013243
• 负责人: Ashutosh Chilkoti
• 金额: $ 38.53万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-01 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10013243
• 关键词: Area; Behavior; Biocompatible Materials; Biology; Biotechnology; Cells; Cholesterol; Development; Disease; Elastin; Exhibits; Genes; Investigation; Length; Libraries; Medicine; Mental Depression; Methods; Modification; Molecular; Peptides; Phase; Phase Transition; Phosphorylation; Polymers; Post-Translational Protein Processing; Property; Proteins; Reaction; Recombinants; Self-Direction; Stimulus; Structural Protein; Structure; Temperature; Work; aqueous; design; heuristics; in vivo; interest; myristoylation; next generation; organizational structure; polypeptide; recombinant peptide; self assembly

Abstract The proposed work in this MIRA application leverages my long-standing interest and expertise in the design of genetically encoded stimulus-responsive peptide polymers. My group pioneered the development of recombinant elastin-like polypeptides (ELPs) that exhibit lower critical solution temperature (LCST) phase behavior. We have also, in parallel, pioneered the development of high-throughput methods for the assembly of highly repetitive genes that we used to create the largest extant library of recombinant peptide polymers. Characterization of their aqueous phase behavior led to the discovery of sequence heuristics that can be used for the de novo used design of peptide polymers that exhibit LCST phase behavior and a class of resilin-like polypeptides (RLPs) that exhibit the converse — upper critical solution phase transition (UCST) phase behavior. Building upon this work, we will explore two new areas in this proposal. First, we will investigate how we can recapitulate the hierarchical structure and properties exhibited by biological materials by the design of partially ordered polymers (POPs) —that consist of disordered polypeptides embedded with a periodically recurring secondary structure motif— that exhibit temperature triggered hierarchical self-assembly into macroscopic materials that mimic the in vivo organization of structural proteins like elastin networks. We will carry out a systematic exploration of the design of new POPs, to verify that the combination of order and disorder at the chain segment level is a new and robust design principle that will yield materials with hierarchical self- assembly across many length scales. Second, we will develop a new line of investigation on genetically encoded biohybrid polymers via post-translational modifications (PTMs) that precisely combine peptide and non-peptide components to create biomaterials that exhibit triggered, hierarchical self-assembly into macroscopic materials. In this aim, we will expand upon our initial work on in vivo myristoylation of ELPs and UCST exhibiting RLPs to investigate if we can convert structure-directing peptides into myristoylation substrates, to create myristoylated polypeptides where the myristoylated segment can direct hierarchical self-assembly of the entire construct. We will also investigate modification of ELPs and RLPs with cholesterol that has the potential to direct self-assembly, and phosphorylation, which will provide a unique trigger of self-assembly. Much remains to be done in both areas, as our preliminary foray into these new areas only hint at the enormous possibilities in the molecular design of new biomaterials enabled by these approaches. The work we propose herein promises to yield new biomaterials with interesting structures and properties with a host of applications in biotechnology and medicine.

摘要 在此MIRA应用程序中的拟议工作利用了我在基因设计方面的长期兴趣和专业知识。 编码的刺激响应肽聚合物。我的小组率先开发了重组弹性蛋白样多肽 在一些实施方案中，所述聚合物是表现出较低临界溶解温度（LCST）相行为的ELP。同时，我们还开创了 开发高通量的方法来组装高度重复的基因，我们用它来创造现存最大的 重组肽聚合物文库。对它们的水相行为的表征导致了序列的发现 可用于从头设计的化学方法使用了表现出LCST相行为和一类具有LCST相行为的肽聚合物。 节枝弹性蛋白样多肽（RLP）表现出匡威上临界溶液相变（UCST）相行为。 在这项工作的基础上，我们将探讨本提案中的两个新领域。首先，我们将研究如何概括 通过部分有序聚合物（POP）的设计， 由嵌入有周期性重复的二级结构基序的无序多肽组成， 引发了分层自组装成宏观材料，模仿体内组织的结构蛋白质，如 弹性蛋白网络我们将对新POP的设计进行系统的探索，验证组合的有序性 链段水平的无序是一种新的稳健的设计原理，它将产生具有分层自 在许多长度尺度上组装。第二，我们将开发一条新的研究路线，对基因编码的生物杂交 聚合物通过翻译后修饰（PTM）精确地将联合收割机肽和非肽组分结合， 生物材料表现出触发，分级自组装成宏观材料。为此，我们将扩大我们的 对ELP和UCST显示RLP的体内豆蔻酰化的初步工作，以研究我们是否可以将结构导向的 肽转化为豆蔻酰化底物，以产生豆蔻酰化的多肽，其中豆蔻酰化的片段可以直接 整个结构的分层自组装。我们还将研究用胆固醇修饰ELP和RLP， 具有指导自组装和磷酸化的潜力，这将提供自组装的独特触发因素。仍有许多工作 在这两个领域都要做，因为我们对这些新领域的初步探索只暗示了分子生物学的巨大可能性。 通过这些方法设计新的生物材料。我们在此提出的工作有望产生新的生物材料， 有趣的结构和性质，在生物技术和医学中具有许多应用。