ProtonsInProteins: A novel approach for studying biological proton transfer

ProtonsInProteins：研究生物质子转移的新方法

• 批准号: EP/Z000459/1
• 负责人: Nadav Amdursky
• 金额: $ 242.91万
• 依托单位: University of Sheffield
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2024
• 资助国家: 英国
• 起止时间: 2024 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=EP%2FZ000459%2F1
• 关键词: ProtonsInProteins; novel; approach; studying; biological

Proton transfer (PT) reactions within proteins are fundamental in biological systems, such as within ATP production. To date, there are no direct means to measure specific PT pathways within proteins, and most research is based on following the end-product of the PT reaction across a natural proton pathway. Here, our goal is to develop a novel way to directly measure PT within proteins. Our new approach is based on placing proton donors and acceptors in specific places within proteins, in which the PT will be initiated only after light excitation of the system. To do so, we introduce here two noncanonical amino acids (ncAA) we developed, with a photoacid and a photobase as their residues that serve as the proton donor and acceptor, respectively. Our hypothesis is that the strong light-triggered driving force of PT in the excited-state (of ~11 pKa units) will initiate PT along the pathway from donor to acceptor. In the first objective, we will use our new ncAA with solid-phase peptide synthesis to design several peptide systems that will allow us to decipher the role of specific amino acids, the peptide structure, and the role of water in PT across the peptide using various ultrafast spectroscopy. In the second objective, we aim to design a mutually orthogonal system for the insertion of our two ncAA into a single protein. In our third objective, we plan to use our new experimental system for answering a unique set of questions in the field of biological PT that could not have been answered before, focusing on the systems of ATP synthase transmembrane complex and the soluble carbonic anhydrase enzyme. Our new approach is groundbreaking in the way we study and understand PT in biology and will enable researchers completely new capabilities resulting in fascinating new discoveries. Moreover, our new system can be translated into other fields in biology that require the local change in proton concentration within proteins.

蛋白质内的质子转移（PT）反应在生物系统中是基本的，例如在ATP生产中。到目前为止，还没有直接的方法来测量蛋白质内的特定PT途径，大多数研究都是基于在天然质子途径上跟踪PT反应的最终产物。在这里，我们的目标是开发一种新的方法来直接测量蛋白质内的PT。我们的新方法是基于将质子供体和受体放置在蛋白质内的特定位置，其中仅在系统的光激发后才会启动PT。要做到这一点，我们在这里介绍两个非规范氨基酸（ncAA），我们开发的，与光酸和光碱作为其残基，作为质子供体和受体，分别。我们的假设是，PT在激发态（约11 pKa单位）的强光触发驱动力将启动PT沿着从供体到受体的途径。在第一个目标中，我们将使用我们的新ncAA与固相肽合成来设计几个肽系统，这将使我们能够使用各种超快光谱来破译特定氨基酸的作用，肽结构以及水在PT中的作用。在第二个目标中，我们的目标是设计一个相互正交的系统，将我们的两个ncAA插入到一个蛋白质中。在我们的第三个目标中，我们计划使用我们的新实验系统来回答生物PT领域中以前无法回答的一组独特问题，重点是ATP合酶跨膜复合物和可溶性碳酸酐酶系统。我们的新方法在生物学中研究和理解PT的方式上具有开创性，并将使研究人员具有全新的能力，从而产生迷人的新发现。此外，我们的新系统可以被翻译到生物学中的其他领域，这些领域需要蛋白质内质子浓度的局部变化。