Expanding the role of P-31 NMR in the study of phosphate-acting enzymes

扩大 P-31 NMR 在磷酸酶研究中的作用

• 批准号: 529394-2018
• 负责人: Berti, Paul
• 金额: $ 1.82万
• 依托单位: McMaster University
• 依托单位国家: 加拿大
• 项目类别: Engage Grants Program
• 财政年份: 2018
• 资助国家: 加拿大
• 起止时间: 2018-01-01 至 2019-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=645340
• 关键词: Expanding; role; 31; NMR; study

Bruker is the world leader in nuclear magnetic resonance (NMR) spectrometers, and is keen to expand its**presence in catalysis and enzyme inhibition research. Many pharmaceutical targets are enzymes that**chemically transform phosphate-containing compounds. Twelve kinase inhibitor drugs are already used in**cancer chemotherapy, and >100 more in are clinical trials. Detailed understanding of enzyme mechanisms**greatly aids new inhibitor design: the flu drugs oseltamivir and zanamivir, and the cancer drug forodesine were**all designed after determining the target enzymes' transition state (TS) structures. The TS is the top of the**energetic barrier that enzymes must surmount for a reaction to occur. Enzymes bind TSs extremely tightly,**making TS mimics potent inhibitors. However, determining TS structures is technically demanding; it requires**measuring the effects of isotopic labels on reaction rates, effects that are frequently < 1%. Recent advances in**NMR spectroscopy have brought the instrumentation for these measurements within reach of most labs, but the**methods needed to exploit these new capabilities remain under-developed. We propose a combined physical**chemistry / applied enzymology study to develop methods to use phosphorus-31 (31P) NMR to measure**isotope effects. This represents an opportune combination of expertise, with Bruker Canada providing the**NMR expertise while the Berti lab providing expertise in TS analysis. Bruker Canada will directly benefit**from the development of new methods that will become part of their applications library. Both private sector**and academic labs will be able to take advantage of the expanded repertoire of NMR techniques to determine**TS structures, expanding the utility of NMR spectroscopy for non-specialist labs. Bruker Canada will also**benefit from direct access to sample preparation, HQP and the NMR spectrometers available at McMaster for**both development and demonstrations for potential customers. The proposed NMR experiments are expected**to attract the interest of other academic laboratories working on inhibitor design and reaction mechanisms. The**proposed work will therefore expand Bruker Canada's customer base in both the private and public sectors.

布鲁克是核磁共振（NMR）光谱仪领域的全球领导者，并热衷于扩大其在催化和酶抑制研究领域的影响力。许多药物靶点是酶，这些酶能使含磷酸盐的化合物发生化学转化。12种激酶抑制剂药物已经用于癌症化疗，超过100种药物正在进行临床试验。对酶机制的详细了解 ** 极大地帮助了新抑制剂的设计：流感药物奥司他韦和扎那米韦，以及癌症药物福罗地辛 ** 都是在确定靶酶的过渡态（TS）结构后设计的。TS是酶发生反应必须克服的能量屏障的顶部。酶与TS的结合非常紧密，** 使TS模拟有效的抑制剂。然而，确定TS结构在技术上要求很高;它需要 ** 测量同位素标记对反应速率的影响，通常<1%。**NMR光谱学的最新进展使大多数实验室都能使用这些测量仪器，但开发这些新功能所需的 ** 方法仍然开发不足。我们提出了一个结合物理 ** 化学/应用酶学研究开发方法，使用磷-31（31 P）NMR测量 ** 同位素效应。这代表了专业知识的适当组合，布鲁克加拿大提供 **NMR专业知识，而Berti实验室提供TS分析专业知识。布鲁克加拿大公司将直接受益于新方法的开发，这些新方法将成为其应用程序库的一部分。私营部门 ** 和学术实验室都将能够利用扩大的NMR技术库来确定 **TS结构，从而扩大NMR光谱学在非专业实验室中的应用。布鲁克加拿大公司还将 ** 受益于直接使用McMaster提供的样品制备、HQP和NMR光谱仪，** 为潜在客户进行开发和演示。建议的核磁共振实验预计 ** 将吸引其他学术实验室的兴趣，抑制剂的设计和反应机理的工作。因此，** 拟议的工作将扩大布鲁克加拿大公司在私营和公共部门的客户群。