Structural and Functional Synthetic Proteomimetics of Ankyrin Repeat Proteins

锚蛋白重复蛋白的结构和功能合成蛋白质模拟

• 批准号: 10537913
• 负责人: Julia Oktawiec
• 金额: $ 6.76万
• 依托单位: NORTHWESTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10537913
• 关键词: Affect; Affinity; Amino Acids; Ankyrin Repeat; Ankyrins; Antibodies; Behavior; Binding Proteins; Biology; CDK4 gene; Cell Cycle; Cell Cycle Progression; Cells; Chemicals; Circular Dichroism Spectroscopy; Clinic; Co-Immunoprecipitations; Complex; Consensus; Cyclin-Dependent Kinase Inhibitor 2A; Development; Differential Scanning Calorimetry; Disease; Disease Progression; Drug Delivery Systems; Drug Targeting; Environment; Enzymatic Biochemistry; Esthesia; Evolution; Fluorescence; Formulation; Goals; Grant; Hydrocarbons; In Vitro; Individual; Inflammatory Response; Investigation; Knowledge; Macular degeneration; Measurement; Mediating; Mentorship; Modality; Molecular Conformation; Mutate; Outcome; Peptides; Pharmaceutical Preparations; Polymer Chemistry; Polymers; Property; Proteins; Research; Research Project Grants; Resources; Roentgen Rays; Structure; Students; Surface; System; Tandem Repeat Sequences; Temperature; Testing; Therapeutic; Thermodynamics; Toxic effect; Training; Tumor Suppressor Proteins; Universities; Vertebral column; Work; Writing; antibody mimetics; base; career; design; experimental study; functional mimics; hydrophilicity; improved; inhibitor therapy; insight; macromolecule; malignant breast neoplasm; mimetics; monomer; peptide drug; polymerization; protein aminoacid sequence; protein protein interaction; scaffold; skills; small molecule; synthetic protein; tool; uptake

With the insight that tandem repeat proteins evolved from more homogeneous ancestors,1–3 our central hypothesis is that their hallmark complex interaction surfaces can be emulated by simple synthetic repeating constructs. We ask two key questions in the proposed research: 1) Can higher order folding of simple, individual ankyrin repeats be induced by spatial organization on synthetic polymeric scaffolds allowing multivalency, and thus replicating such interaction surfaces? and 2) What is the simplest ankyrin repeat that can functionally recapitulate a high affinity PPI? To probe these questions, a polymer-based approach to proteomimetics will be employed. The proposed proteomimetic systems will provide insight into ankyrin repeat proteins by emulating the properties of proteins while remaining synthetically straightforward and structurally simple.4 Specifically, we propose protein-like polymers (PLPs)5–10 as these constructs are monodisperse, protein- sized macromolecules that are rapidly and scalably produced by polymerization of peptide-based monomers. The result is a peptide brush polymer wherein peptides are tethered to a hydrocarbon polymer backbone in a dense display. This results in a peptide topology that resembles repeat proteins, but where the peptides repeats are arranged on a synthetic scaffold.6,7 We hypothesize that this proteomimetic platform can enable conformational mimics of ankyrin repeat proteins, to serve as effective mimics for the disruption of PPIs as tools and therapeutics. The long-term goal of this application is the precise design of tandem repeat-mimetic PLPs for the formulation and delivery of therapeutic peptides and as chemical biology tools. With the combined knowledge from these lines of inquiry, reliable rules will be established for the creation of highly stable, yet effective PLPs that target protein-protein interactions in diseases. Furthermore, this knowledge will inform the design of proteomimetic materials for therapeutic applications more broadly. In addition to this research project, a training plan will be pursued to develop the expertise and proficiency of the candidate in soft matter synthesis and characterization, drawing upon the collaborative and motivated research environment in Prof. Gianneschi's group and the considerable resources available through Northwestern University. This plan will include honing skills in mentorship of students, presenting results, effective grant-writing, and the management of research groups. This training plan and research environment will help the candidate grow in their transition to new fields and lines of inquiry, towards the pursuit of an independent research career investigating bio-inspired materials for targeted drug delivery.

随着串联重复序列蛋白质从更同质的祖先进化而来的见解，1-3我们的中心 假设是，它们的标志性复杂相互作用表面可以通过简单的合成来模拟。 重复构建。我们提出了两个关键问题，在拟议的研究：1）可以高阶折叠的简单， 通过在合成聚合物支架上的空间组织诱导单个锚蛋白重复， 多价，从而复制这种相互作用的表面？和2）什么是最简单的锚蛋白重复，可以 在功能上概括了高亲和力PPI？为了探讨这些问题，一种基于聚合物的方法， 将使用蛋白质模拟物。所提出的蛋白质模拟系统将提供深入了解锚蛋白重复序列 通过模拟蛋白质的性质，同时保持合成简单和结构 简单。4具体地说，我们提出了蛋白质样聚合物（PLPs）5-10，因为这些构建体是单分散的，蛋白质- 通过基于肽的单体的聚合快速且可规模化地产生的尺寸大分子。 结果是肽刷状聚合物，其中肽以聚合物链的形式连接到烃聚合物主链上。 密集显示。这导致了类似于重复蛋白质的肽拓扑结构，但其中肽重复 排列在合成支架上。6，7我们假设这种蛋白质模拟平台可以使 锚蛋白重复序列蛋白的构象模拟物，作为破坏PPI的有效模拟物， 和治疗学。本申请的长期目标是精确设计串联重复序列模拟PLPs， 治疗性肽的配制和递送以及作为化学生物学工具。有了综合的知识 从这些调查线索中，将建立可靠的规则，以创建高度稳定但有效的PLP 针对疾病中蛋白质间的相互作用。此外，这些知识将告知设计 蛋白质模拟材料用于更广泛的治疗应用。 除了这个研究项目外，还将实施一项培训计划， 候选人在软物质合成和表征，借鉴合作和积极的 Gianneschi教授小组的研究环境以及通过 西北大学。该计划将包括磨练学生导师的技能，展示成果， 有效的赠款写作和研究小组的管理。这种培训计划和研究环境 将帮助候选人在过渡到新的领域和调查路线中成长， 独立的研究生涯，调查生物启发材料的靶向药物输送。