Molecular Mimics of Protein Tertiary Folding from Primary Sequence Information

项目总结 蛋白质是使生命得以存在的核心功能工具，并为 蛋白质模拟对化学家来说是一个巨大的挑战。有明确折叠倾向的人造脊椎， 被称为“Folamers”，可以提供自然实体的生物稳定的类似物；然而，与设计相关的挑战 为模仿复杂的三级折叠设置障碍。克服这一障碍有望开辟一条新的前沿 并使折叠剂朝着蛋白质的功能多样性方向发展。 在最初奖项的支持下，创建更具折叠式三级结构的一般方法是 基于天然序列中主链共价结构的系统变化而发展起来的。一个 在建立这些模拟物复制和调节功能的能力方面仍然存在重要的差距 它们所基于的原型蛋白质的性质。国际和平研究所研究计划的一个长期目标是 开发能够复制完整蛋白质的人工脊椎的设计原则 并将这些原理应用于控制诸如折叠结构的性质， 折叠稳定性、生理稳定性和动力学。此续期申请的总体目标是 演示异类主干折叠式三级结构中可能的功能范围 模仿学。指导这项工作的中心假设是在最初的奖项中制定的设计原则 Period可以用来产生不同原型蛋白的功能类似物，也可以用来调整 本机主干的功能特征。进行这项拟议研究的理由是 在以蛋白质为灵感的人工支架中推动超越结构模仿到功能模仿将会磨练 设计原则，创造有价值的生物活性物质，并为自然系统提供新的认识。 为了验证上述中心假设，将追求两个具体目标：(1)开发模拟 锌指蛋白具有天然的分子识别特性；(2)产生富含二硫键的模拟物 来自昆虫和爬行动物毒液的区域。 在预期成果方面，拟议的工作将(1)扩大高等教育的范围 结构拟态(复杂的链状拓扑结构，大的多结构域蛋白质)；(2)拓宽功能 这些支架的系列(选择性识别DNA、蛋白质和生物膜)；(3)产率 对锌指蛋白序列特异性DNA结合动力学的新见解；以及(4)提供了一种 生物活性药物在慢性疼痛和疼痛治疗中的潜在应用 微生物感染的治疗。总体而言，项目目标的实现将导致垂直 在合成蛋白质模拟中可能的大小、结构复杂性和功能多样性方面的进展。

PROJECT SUMMARY Proteins are the central functional instruments that enable life, and the development of strategies for protein mimicry is a grand challenge for chemists. Artificial backbones with defined folding propensities, termed “foldamers”, can offer biostable analogues of natural entities; however, challenges related to design create barriers to mimicking complex tertiary folds. Overcoming this barrier promises to open a new frontier and advance foldamers toward the functional versatility of proteins. With support from the initial award, a general method for creating foldamer tertiary structure was developed based on the systematic alteration of backbone covalent structure in natural sequences. An important gap remains in establishing the ability of these mimetics to reproduce and modulate functional properties of prototype proteins on which they are based. A long-term goal of the PI’s research program is to develop principles for the design of artificial backbones capable of reproducing the full panoply of protein folds and functions in nature and to apply these principles to control properties such as folded structure, folded stability, physiological stability, and dynamics. The overall objective of this renewal application is to demonstrate the scope of functions possible in heterogeneous-backbone foldamer tertiary structure mimetics. The central hypothesis guiding this work is that design principles developed in the initial award period can be applied to produce functional analogues of diverse prototype proteins and also used to tune functional characteristics of the native backbone. The rationale for pursuing the proposed research is that pushing beyond structural mimicry to functional mimicry in protein-inspired artificial scaffolds will hone design principles, create valuable bioactive agents, and shed new light on natural systems. In order to test the above central hypothesis, two specific aims will be pursued: (1) develop mimics of zinc finger proteins with native-like molecular recognition characteristics; (2) create mimics of disulfide-rich domains from insect and reptile venoms. In terms of expected outcomes, the proposed work will (1) expand the scope of foldamer tertiary structure mimicry (complex chain topologies, large multidomain proteins); (2) broaden the functional repertoire of these scaffolds (selective recognition of DNA, proteins, and biological membranes); (3) yield new insights into the dynamics of sequence-specific DNA binding by zinc finger proteins; and (4) provide a starting point toward bioactive agents with potential applications in the management of chronic pain and treatment of microbial infections. Collectively, realization of the goals of the project will lead to a vertical advance in the size, structural complexity, and functional diversity possible in synthetic protein mimetics.