Genetically Encoded Smart Biohybrid Materials

摘要 这个Mira应用程序中建议的工作利用了我在基因设计方面的长期兴趣和专业知识 编码的刺激反应性多肽聚合物。我的团队率先开发了重组类弹性蛋白多肽 (ELPS)表现出较低的临界溶液温度(LCST)相行为。同时，我们还开创了 开发高通量的方法来组装我们用来创造最大现存基因的高度重复的基因 重组多肽聚合物库。对它们的水相行为的表征导致了序列的发现 启发式算法，可用于从头设计呈现LCST相行为的多肽聚合物和一类 Resilin样多肽(RLP)，表现出逆-上临界溶液相变(UCST)相行为。 在这项工作的基础上，我们将在这项提议中探索两个新领域。首先，我们将研究如何重述 生物材料通过偏序聚合物(POP)的设计表现出的层次结构和性质-- 由嵌有周期性重复的二级结构基序的无序多肽组成，显示出温度 触发分层自组装成宏观材料，模拟结构蛋白的体内组织，如 弹性蛋白网络。我们将对新POP的设计进行系统的探索，以验证顺序的结合 而链段水平的无序是一种新的、健壮的设计原则，它将产生具有分层自组织结构的材料。 跨多个长度刻度的装配。第二，我们将开发一条新的遗传编码生物杂交研究路线 通过翻译后修饰(PTM)精确结合多肽和非肽成分的聚合物 生物材料表现出触发的、层次化的自组装成宏观材料。在这一目标中，我们将扩展我们的 显示RLP的ELPs和UCST的体内肉豆蔻酰化的初步工作，以研究我们是否可以转换结构导向 多肽进入肉豆蔻化底物，以产生肉豆蔻化多肽，在肉豆蔻化片段可以定向 整个构造的分层自组装。我们还将研究用胆固醇修饰ELP和RLP 具有直接自组装和磷酸化的潜力，这将提供一个独特的自组装触发。剩下的很多 在这两个领域都要完成，因为我们对这些新领域的初步尝试只暗示了分子中的巨大可能性 通过这些方法设计新的生物材料。我们在这里提出的工作承诺用来生产新的生物材料 有趣的结构和性质以及在生物技术和医学中的大量应用。

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.