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.

核磁共振波谱是研究分子结构、动力学和化学键的一种强大的分析技术。这项技术在过去几十年的发展使化学和生物科学发生了革命性的变化。然而，大多数成功的生物体系的核磁共振研究，如蛋白质和核酸，主要是基于检测核自旋量子数为1/2的原子核，如1H，13C和15N。另一方面，氧元素作为有机和生物分子中含量最丰富的元素之一，还没有被核磁共振波谱所获得。这主要是因为唯一的核磁共振活性氧同位素17O的核自旋量子数为5/2(称为四极核)。此外，17O的自然丰度极低，为0.037%。众所周知，四极核很难用核磁共振进行研究，因为它们通常会产生非常广泛的核磁共振信号。因此，在生物大分子的背景下，17O常常被认为是“核磁共振不可见的”。最近，我们在解决这一具有挑战性的问题的道路上取得了两项突破。首先，我们开发了新的核磁共振方法，可以检测固体和水溶液中生物大分子的高分辨率17O核磁共振谱。第二，我们用营养缺陷型大肠杆菌菌株展示了一种将17O整合到重组蛋白中的一般方法。在接下来的5年里，我们将把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核磁共振谱通常被认为是生物分子核磁共振的最后一个前沿。我们将探索17O核磁共振在蛋白质结构和动力学研究中所能提供的独特信息。我们将研究17O核磁共振作为一种新的示踪剂用于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核磁共振作为一种补充工具，可能对癌症诊断和监测有用。我们研究计划的最终目标是使在生物分子中发现的所有主要元素(H、C、N和O)都可以通过核磁共振光谱学获得。在这一领域，我们的研究项目一直处于国际领先地位。拟议研究的结果将对化学和生物科学的许多学科产生影响。拟议的研究计划还将为培养具有全球竞争力的下一代高素质人才提供极好的机会。