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; 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.

有了以下洞察力，即串联重复蛋白从更均匀的祖先演变而来，1-3我们的中央 假设是可以通过简单的合成来模拟它们的Hallmark复合体相互作用表面 重复构造。我们在拟议的研究中提出了两个关键问题：1）可以更高阶折叠简单， 空间组织在合成聚合物支架上诱导单个伸缩蛋白重复。 多价，从而复制此类相互作用表面？ 2）什么是最简单的氨基林重复 在功能上概括了高亲和力PPI？为了探究这些问题，一种基于聚合物的方法 将雇用蛋白质组合。所提出的蛋白质组合系统将提供深入了解arnkyrin的重复 蛋白质通过模拟蛋白质的特性，同时保持合成直接和结构上的蛋白质 简单4，具体，我们提出了蛋白质样聚合物（PLP）5-10，因为这些构建体是单分散的，蛋白质 - 大小的大分子是通过基于肽的单体聚合而迅速且可扩展产生的。 结果是辣椒刷聚合物，将辣椒绑在碳氢化合物聚合物主链上 密集显示。这会导致类似于重复蛋白的肽拓扑，但是肽重复 安排在合成支架上。6,7我们假设这个蛋白质映射平台可以启用 脚踝重复蛋白的构象模仿，可以作为破坏PPI作为工具的有效模仿 和治疗。该应用的长期目标是串联重复模拟PLP的精确设计 热毛皮的配方和传递作为化学生物学工具。与合并知识 从这些探究方面，将建立可靠的规则，以创建高度稳定但有效的PLP 该靶向疾病中的蛋白质蛋白质相互作用。此外，这些知识将为设计 用于治疗应用的蛋白质材料更广泛。 除了该研究项目外，还将采用培训计划来发展专业知识和熟练程度 候选人在软物质综合和表征中的依靠协作和成熟的 Gianneschi教授的研究环境以及可通过的大量资源 西北大学。该计划将包括磨练学生指导的技能，提出结果， 有效的赠款写作和研究小组的管理。这个培训计划和研究环境 将帮助候选人在过渡到新领域和探究方面成长，以追求 独立的研究职业研究以生物启发的材料进行有针对性的药物输送。