Development of new 17O NMR spectroscopic techniques for studying biological systems

开发用于研究生物系统的新型 17O NMR 波谱技术

• 批准号: RGPIN-2021-03140
• 负责人: Wu, Gang
• 金额: $ 5.76万
• 依托单位: Queen's University
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2022
• 资助国家: 加拿大
• 起止时间: 2022-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=748163
• 关键词: Development; new; 17O; NMR; spectroscopic

Nuclear magnetic resonance (NMR) spectroscopy is a powerful analytical technique for studying molecular structure, dynamics, and chemical bonding. The development of this technique over the past several decades has revolutionized chemical and biological sciences. However, most successful NMR studies of biological systems such as proteins and nucleic acids are primarily based on detection of atomic nuclei with a nuclear spin quantum number of 1/2 such as 1H, 13C, and 15N. On the other hand, the oxygen element, being one of the most abundant in organic and biological molecules, has not yet been accessible by NMR spectroscopy. This is principally because the only NMR-active oxygen isotope, 17O, has a nuclear spin quantum number of 5/2 (known as a quadrupolar nucleus). In addition, the natural abundance of 17O is extremely low, 0.037%. Quadrupolar nuclei are notoriously difficult to study by NMR because they often give rise to very broad NMR signals. As a result, 17O is very often considered to be "NMR invisible" in the context of biological macromolecules. Recently, we made two breakthroughs on the way of solving this challenging problem. First, we have developed new NMR methods that allow detection of high-resolution 17O NMR spectra for biological macromolecules both in the solid state and in aqueous solution. Second, we have demonstrated a general method of incorporating 17O into recombinant proteins with auxotrophic E. coli strains. In the next 5 years, we will apply these new methods to two types of biological systems: proteins and live cancer cells. 17O NMR spectroscopy of proteins is often considered to be the last frontier of biomolecular NMR. We will explore the unique information that 17O NMR can offer in the study of protein structure and dynamics. We will investigate the use of 17O NMR as a new tracer for monitoring glucose and glutamine metabolism in live cancer cells. This will help establish 17O NMR as a complementary tool potentially useful for cancer diagnosis and monitoring. The ultimate goal of our research program is to make all major elements (H, C, N, and O) found in biological molecules equally accessible by NMR spectroscopy. In this field, our research program has been an international leader. The outcomes from the proposed research will have impact in many disciplines in chemical and biological sciences. The proposed research program will also provide excellent opportunities for training of the next generation of highly qualified personnel to be competitive globally.

核磁共振（NMR）光谱是一种用于研究分子结构，动力学和化学键合的强大分析技术。在过去的几十年中，这种技术的发展彻底改变了化学和生物科学。但是，大多数成功的NMR研究（例如蛋白质和核酸）主要基于对原子核的检测，其核自旋量子数为1/2，例如1H，13C和15N。另一方面，NMR光谱尚未访问有机和生物分子中最丰富的氧元素之一。这主要是因为唯一的NMR活性氧同位素为17O，其核自旋量子数为5/2（称为四核核）。另外，17o的自然丰度极低，0.037％。众所周知，NMR很难研究四极核，因为它们通常会产生非常广泛的NMR信号。结果，在生物大分子的背景下，经常认为17o被认为是“ NMR不可见的”。最近，我们在解决这个具有挑战性的问题的方式上取得了两个突破。首先，我们开发了新的NMR方法，该方法允许检测高分辨率17O NMR光谱，用于固态和水溶液中的生物大分子。其次，我们已经证明了一种将17O掺入具有补充营养大肠杆菌菌株的重组蛋白中的一般方法。在接下来的5年中，我们将将这些新方法应用于两种类型的生物系统：蛋白质和活癌细胞。蛋白质的17O NMR光谱通常被认为是生物分子NMR的最后边界。我们将探讨17o NMR在蛋白质结构和动力学研究中可以提供的独特信息。我们将研究17o NMR作为新的示踪剂，以监测活癌细胞中的葡萄糖和谷氨酰胺代谢。这将有助于建立17o NMR作为一种互补工具，可用于癌症诊断和监测。我们的研究计划的最终目标是使在NMR光谱法中同样访问的生物分子中发现所有主要元素（H，C，N和O）。在这个领域，我们的研究计划一直是国际领导者。拟议的研究的结果将影响化学和生物科学的许多学科。拟议的研究计划还将为培训下一代高素质的人员提供极好的机会，以使其在全球范围内具有竞争力。