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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

基本信息

批准号：
10618928
负责人：
Brett VanVeller
金额：
$ 36.5万
依托单位：
IOWA STATE UNIVERSITY
依托单位国家：
美国
项目类别：
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

项目摘要

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/摘要本申请的目的是开发立体化学健壮的硫代酰胺，在多肽Se-Se中的任何一点- QUENCE，作为生物物理探测器，以解决目前在肽合成、折叠和药物发现方面的障碍。人们认为，目前将硫代酰胺结合到多肽中的方法是足够的。然而，由于例如，在合成过程中硫酰胺残基的α-碳立体化学的快速消旋掩盖了这种看法。事实上，一项对报道的含有硫代酰胺的多肽的调查指出了这些局限性。大多数成功的序列都将硫代酰胺结合到N-末端，在那里暴露于在Fmoc固相肽合成(SPPS)过程中，合成试剂必须最小化。因此，硫代酰胺在Fmoc SPPS过程中的不稳定性是合成和评价硫代酰胺的重要障碍。硫代酰胺多肽的功能，限制了硫代酰胺类化合物的序列空间。轶事报告表明，许多实验室都希望在各种多肽研究中使用硫代酰胺，但缺乏记录在案的陷阱和合成选择导致难以处理的多肽产品，并最终被丢弃这样的企业。这项建议的方法是保护硫代酰胺，类似于保护氨基酸侧链的官能团，以便在肽延伸过程中保持硫代酰胺部分。提顿。这种方法的基本原理是硫代酰胺保护可以很容易地包括在标准中 SPPS工作流程使其能够在多肽合成、骨架修饰和蛋白质药物中实现新的应用互动。该项目的研究计划将利用硫代酰胺来探测蛋白质折叠和位点选择性插入其他化学物质。将在以前未知的序列空间中使用硫代酰胺来寻址蛋白质折叠中的基本问题。我们还将开发将硫代酰胺转化为功能性的方法将解锁新的受限制的多肽支架的团体。具有持久结构的多肽具有巨大的承诺作为治疗药物，弥合小分子和生物制品之间的性能差距。最后，这一点工作将确定询问和定位与治疗相关的蛋白质-蛋白质界面的策略。Interac- 多肽主链氢键供体和受体与蛋白质结合靶标的相互作用在药物设计中没有得到充分利用。基于结构生物信息学，识别未充分利用的新战略-- 蛋白质-蛋白质界面(PPI)的相互作用将有助于设计更有效的抑制剂。其他工作将还要开发新的工具来询问PPI，这些PPI的结构信息可能非常少未来的实验设计。这项拟议的研究具有创新性，因为它代表了一项实质性的突破从现状出发，开发和采用新的方法来保持硫代酰胺的稳定性，这有望开启新的研究视野。这一贡献意义重大，因为预计它将产生广泛的影响。在蛋白质折叠的研究和生物活性分子的开发方面都具有重要意义。