Peptide backbone modifications to enhance and study protein folding and binding

项目概要/摘要 本申请的目的是为一项对执行《公约》至关重要的文书获得补充资金。 NIGMS资助R35 GM 142883的研究目标。支持的研究旨在利用立体化学- 在肽序列中的任何点上，作为生物物理探针来解决电流障碍 在肽合成、折叠和药物发现方面。该提议中的方法是保护硫代酰胺， 类似于保护氨基酸侧链的官能团，以保留硫代酰胺 在肽延伸期间的部分。这种方法的基本原理是硫代酰胺保护可以很容易地在- 包括在标准SPPS工作流程中，以实现肽合成、骨架修饰、 阳离子和蛋白质-药物相互作用。本项目的研究计划是利用硫代酰胺类化合物作为蛋白质探针 折叠和位点选择性插入等化学反应。硫代酰胺将用于以前未知的领域， 序列空间来解决蛋白质折叠的基本问题。我们还将开发方法， 将硫代酰胺转化为官能团，这将解锁新的受限肽支架。具有持久性的肽 结构作为治疗剂具有巨大的希望，可以弥合小分子之间的性能差距， 和生物制剂。最后，这项工作将确定询问和靶向治疗相关蛋白质的策略， 蛋白质界面多肽主链上氢键供体与受体之间的相互作用 和蛋白质结合靶点在药物设计中未得到充分利用。基于结构生物信息学， 确定蛋白质-蛋白质界面（PPI）未充分利用的相互作用将有助于设计更有效的 抑制剂的其他工作还将开发新的工具来询问结构信息很少的PPI 可用于指导未来的实验设计。这项研究是创新的，因为它代表了- 通过开发和采用新的方法来保护硫， 酰胺稳定性，这有望开启新的研究视野。贡献是巨大的，因为它是 预期在蛋白质折叠的研究和生物活性分子的开发中具有广泛的重要性， 克里斯本申请中描述的仪器将支持上述所有活动， 完成所需的。

PROJECT SUMMARY/ABSTRACT The objective in this application is to obtain supplemental funding for an instrument that is vital to carry out the research objectives of NIGMS grant R35 GM142883. The supported research seeks to exploit stereochemi- cally robust thioamides, at any point in the peptide sequence, as biophysical probes to address current barriers in peptide synthesis, folding, and drug discovery. 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 elongation. The rationale for this approach is that thioamide protection can be easily in- cluded within the standard SPPS work-flow to enable novel applications in peptide synthesis, backbone modifi- cation, 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 se- quence 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. Interactions 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 interactions 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 repre- sents a substantive departure from the status quo by developing and employing new methods to preserve thio- amide stability, which promises to unlock new research horizons. The contribution is significant because it is expected to have broad importance in both the study of protein folding and the development of bioactive mole- cules. The instrumentation described in this application will support all of the activities outlined above and is necessary for their completion.