Peptide backbone modifications to enhance and study protein folding and binding

肽骨架修饰以增强和研究蛋白质折叠和结合

• 批准号: 10435444
• 负责人: Brett VanVeller
• 金额: $ 36.57万
• 依托单位: IOWA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10435444
• 关键词: Address; Amino Acids; Basic Science; Binding Proteins; Bioinformatics; Biological Products; Biophysics; Carbon; Chemistry; Development; Diagnosis; Drug Design; Drug Interactions; Evaluation; Experimental Designs; Exposure to; Future; Hydrogen Bonding; Methods; Mission; Modification; Peptide Synthesis; Peptides; Perception; Performance; Phase; Prevention; Procedures; Proteins; Public Health; Reagent; Reporting; Research; Side; Site; Solid; Structure; Surveys; Therapeutic; Therapeutic Intervention; Thioamides; United States National Institutes of Health; Vertebral column; Work; base; design; drug discovery; functional group; human disease; inhibitor; innovation; knowledge base; novel; preservation; programs; protein folding; protein function; racemization; scaffold; small molecule; stereochemistry; targeted treatment; tool

PROJECT SUMMARY/ABSTRACT The objective in this application is to exploit stereochemically robust thioamides, at any point in the peptide se- quence, as biophysical probes to address current barriers in peptide synthesis, folding, and drug discovery. There is a perception that current methods to incorporate thioamides into peptides are sufficient. However, as an example, the rapid racemization of the alpha-carbon stereochemistry of thioamide residues during synthesis belies that perception. Indeed, a survey of reported peptides containing thioamides points to these limitations. The majority of ‘successful’ sequences incorporate the thioamide close to the N-terminus, where exposure to synthetic reagents is necessarily minimized during Fmoc solid-phase peptide synthesis (SPPS) procedures. Thus, the instability of thioamides during Fmoc SPPS present a significant barrier to the synthesis and evalua- tion of thioamide peptides, and restricts the sequence space in which thioamides can be employed. Anecdotal reports indicate that many labs have wished to employ thioamides in a variety of peptide studies, but a lack of documented pitfalls and synthetic options leads to intractable peptide products and, ultimately, abandonment of such ventures. The approach in this proposal is to protect the thioamide, in analogy to the protection of the functional groups of amino acid side chains, in order to preserve the thioamide moiety during peptide elonga- tion. The rationale for this approach is that thioamide protection can be easily included within the standard SPPS work-flow to enable novel applications in peptide synthesis, backbone modification, and protein-drug interactions. The research plans of this project will exploit thioamides to probe protein folding and site-selective insertion other chemistries. Thioamides will be employed in previously uncharted sequence space to address fundamental questions in protein folding. We will also develop methods to transform thioamides into functional groups that will unlock new constrained peptide scaffolds. Peptides with persistent structure hold tremendous promise as therapeutics to bridge the performance gap between small molecules and biologics. Finally, this work will identify strategies to interrogate and target therapeutically relevant protein-protein interfaces. Interac- tions between hydrogen bond donors and acceptors of the main-chain of a peptide and a protein binding target are underutilized in drug design. Based on structural bioinformatics, new strategies to identify underutilized in- teractions at protein-protein interfaces (PPIs) will assist in the design of more potent inhibitors. Other work will also develop new tools to interrogate PPIs for which very little structural information may be available to inform future experimental design. The proposed research is innovative because it represents a substantive departure from the status quo by developing and employing new methods to preserve thioamide stability, which promises to unlock new research horizons. The contribution is significant because it is expected to have broad im- portance in both the study of protein folding and the development of bioactive molecules.

项目摘要/摘要 该应用的目的是在辣椒se的任何时刻探索立体化学上强大的硫酰胺 示例，作为解决肽合成，折叠和药物发现中当前障碍的生物物理问题。 人们认为，将硫酰胺掺入肽中的当前方法就足够了。但是，如 一个例子，在合成过程中，硫酰胺的α-碳立体化学的快速种族化保留 掩盖了那些感知的。实际上，对含有硫酰胺的Petides进行的一项调查表明，这些局限性。 大多数“成功”序列均包含靠近N末端的硫酰胺，其中暴露于 在FMOC固相肽合成（SPPS）过程中，合成试剂必然会最小化。 这就是FMOC属于硫酰胺的不稳定性，这是合成和评估的显着障碍 硫酰胺肽的影响，并限制了可以雇用硫酰胺的序列空间。轶事 报告表明，许多实验室希望在各种肽研究中使用硫酰胺，但缺乏 有记录的陷阱和合成选项导致顽固的肽产品，并最终放弃 这样的冒险。该提议中的方法是保护硫酰胺，类似于保护 氨基酸侧链的官能团，以保留胡椒弹性期间的硫酰胺部分 tion。这种方法的理由是可以轻松将硫酰胺保护包括在标准中 spps的工作流以实现肽合成，骨干修饰和蛋白质 - 药物的新应用 互动。该项目的研究计划将利用硫酰胺探测蛋白质折叠和现场选择性 插入其他化学。硫酰胺将在先前未知的序列空间中雇用以解决 蛋白质折叠中的基本问题。我们还将开发将硫酰胺转化为功能的方法 将解锁新约束肽支架的组。具有持久结构的肽可持续 有望作为弥合小分子与生物制剂之间的性能差距的疗法。最后，这个 工作将确定询问和靶向热相关蛋白质 - 蛋白质界面的策略。间 氢键供体和肽主链受体之间的影响与蛋白质结合靶 在药物设计中不足。基于结构性生物信息学，确定未利用的内部策略 蛋白质 - 蛋白质界面（PPI）的畸形将有助于设计更多的潜在抑制剂。其他工作将 还开发了新工具来询问PPI，几乎没有结构信息可以告知其结构信息 未来的实验设计。拟议的研究具有创新性，因为它代表了实质性的离开 通过开发和采用新方法来维护硫酰胺稳定性，从现状开始，这承诺 解锁新的研究视野。这项贡献很重要，因为预计它将具有广泛的影响 在蛋白质折叠的研究和生物活性分子的发展方面的一部分。