Ultra-low-temperature (6 K) static NMR-DNP for metalloproteins, proteins in cells, and materials

用于金属蛋白、细胞中蛋白质和材料的超低温 (6 K) 静态 NMR-DNP

• 批准号: 10546201
• 负责人: Francis DAVID Doty
• 金额: $ 29.96万
• 依托单位: DOTY SCIENTIFIC, INC.
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-03-01 至 2024-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10546201
• 关键词: Alzheimer' s Disease; Amyloid Fibrils; Antibiotics; Area; Awareness; Bee Venoms; Binding; Biological; Biomedical Research; Budgets; Cells; Cellular Membrane; Country; DNA Repair Gene; Data; Development; Electrons; Equipment; Extremely High Frequency Radio Waves; Health; Human; Inflammasome; Laboratories; Lipids; Magic; Magnetism; Measures; Membrane Proteins; Metalloproteins; Methods; Molecular; Motion; Nisin; Nuclear; Nuclear Magnetic Resonance; Optics; Peptides; Pharmacologic Substance; Phase; Play; Price; Proline; Proteins; Relaxation; Reporting; Research; Research Personnel; Resolution; Role; Sampling; Signal Transduction; Small Business Innovation Research Grant; Solid; Source; Structure; System; Techniques; Temperature; amyloid structure; cold temperature; cost; design; enhancing factor; experimental study; improved; in silico; instrumentation; macromolecule; materials science; microwave electromagnetic radiation; novel; peptide structure; prototype; restraint; simulation; solid state; solid state nuclear magnetic resonance; structural biology; tool; waveguide

Ultra-low-temperature (6 K) static NMR-DNP for metalloproteins, proteins in cells, and materials Abstract The critical importance of solid-state NMR (ssNMR) was recently demonstrated by, after nearly two decades of intense efforts, yielding the first atomic-resolution structures of the A40 and A42 amyloid fibrils that play a crucial role in Alzheimer’s Disease (AD). Challenges posed by the inherently low sensitivity of NMR can be improved by reducing the sample and circuit temperature to below 100 K, or preferably below 35 K – also known as Ultra Low Temperature (ULT). Combining ULT NMR with another emerging technique, denoted as dynamic nuclear polarization (DNP), shows enormous promise for expanding the role of ssNMR by providing significant S/N enhancements in many cases. However, despite the huge gain in S/N that is possible from DNP, also performed at ~90 K, there are still only a handful of high-field MAS-DNP systems in the U.S. – pri- marily because they are so expensive ($3-10M). Further, no commercially available instrumentation is availa- ble for NMR experiments below 90 K, let alone ULT experiments combined with DNP. A huge step forward would include the development of a high-mode THz cavity for more efficient microwave delivery to the sample, thus enabling the use of a lower power and lower cost microwave source. An area of ssNMR that employs the use of non-spinning (static) samples would benefit immensely from the availability of such a commercially built ULT probe, with option for inclusion of DNP. Further, the static probe can serve as a prototype for a MAS ver- sion of such a probe. The developments proposed under this project aim to reduce entry level cost into DNP by more than an order of magnitude while reducing certain operational challenges.

用于金属蛋白、细胞中蛋白质和材料的超低温 (6 K) 静态 NMR-DNP 抽象的 经过近二十年的研究，最近证明了固态核磁共振 (ssNMR) 的至关重要性 经过不懈的努力，产生了 A40 和 A42 淀粉样原纤维的第一个原子分辨率结构，这些结构在 NMR 固有的低灵敏度带来了挑战。 通过将样品和电路温度降低至 100 K 以下，或最好低于 35 K 来改善 – 也 称为超低温 (ULT)，将 ULT NMR 与另一种新兴技术相结合，表示为 动态核极化 (DNP) 通过提供 在许多情况下信噪比显着增强 然而，尽管信噪比可以得到巨大的提高。 DNP，也在 ~90 K 下执行，美国仍然只有少数高场 MAS-DNP 系统 – pri- 主要是因为它们非常昂贵（3-1000万美元），此外，还没有商用仪器可用。 可以用于低于 90 K 的 NMR 实验，更不用说与 DNP 结合的 ULT 实验了。 将包括开发高模太赫兹腔，以更有效地将微波传输到样品， 从而能够使用更低功率和更低成本的微波源。 使用非旋转（静态）样品将极大地受益于这种商业化构建的可用性 ULT 探头，可选择包含 DNP 此外，静态探头可以用作 MAS 版本的原型。 该项目提出的开发旨在降低 DNP 的入门成本。 提高了一个数量级以上，同时减少了某些运营挑战。