An HXYZ-g HR-Fast-MAS probe for Dramatically Improved Biomolecular Structure Determinations

用于显着改进生物分子结构测定的 HXYZ-g HR-Fast-MAS 探针

• 批准号: 9988618
• 负责人: Francis DAVID Doty
• 金额: $ 64.31万
• 依托单位: DOTY SCIENTIFIC, INC.
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-03-01 至 2020-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9988618
• 关键词: Alzheimer' s Disease; Amyloid beta-Protein; Area; Biological; Biomedical Research; Budgets; COSY; Caliber; Catalysis; Cellular Membrane; Complex; Data; Deposition; Detection; Development; Diffusion; Equipment; Funding; Goals; Health; Inferior; Laboratories; Liquid substance; Magic; Magnetism; Measurement; Membrane Proteins; Metabolism; Methods; Modeling; Molecular; Molecular Weight; Motion; NMR Spectroscopy; Neurology; Nuclear; Nuclear Magnetic Resonance; Performance; Phase; Physiologic pulse; Platelet Factor 4; Price; Proteins; Protocols documentation; Publishing; RF coil; Research Personnel; Resolution; Sampling; Scheme; Side; Solid; Solvents; Sonication; Speed; Structural Protein; Structure; Surface; System; Techniques; Technology; Time; Width; base; data warehouse; design; experimental study; improved; innovation; insight; interest; irradiation; macromolecule; materials science; microwave electromagnetic radiation; next generation; novel; operation; protein structure; solid state nuclear magnetic resonance; structural biology; tool; vibration; virtual

An HXYZ-g HR-Fast-MAS probe for Dramatically Improved Biomolecular Structure Determinations Abstract. Liquid state NMR spectroscopy is arguably one of the best tools for structure determination for soluble proteins. The method provides atomic resolution for modest molecular weight proteins and/or their complexes. The method begins to have difficulty when the molecular weight of the system causes slow molecular motion, which in turn increases the linewidth beyond the point of useful resolution. Solid state NMR (ssNMR) methods have progressed remarkably over the past 15 years to permit improved resolution for these conditions, but they still come well short of the goal of liquid-like resolution on biological macromolecules, such as membrane proteins and the fibrils that are central to Alzheimer’s Disease. The “Holy Grail” in ssNMR would be the ability to successfully utilize the powerful suite of NMR acquisition and automated structure determination protocols developed for solution NMR, which rely on ¹H-detected triple- and quad-resonance ²H-decoupled schemes (as such generally permit 8 or 30 times higher S/N than direct detection, for ¹³C and¹⁵5N respectively) with solid samples of 1-10 mg. The main objective of this Phase II application is to complete the development a four-channel multinuclear ssNMR probe (HXYZ) that has the capability of providing ¹³C/¹⁵N/¹H correlations under ²H decoupling utilizing modest (15 kHz) to fast (> 35 kHz) Magic Angle Spinning (MAS) while detecting ¹H. The resulting resolution, particularly with proposed novel pulse sequences, will be close to that of a typical liquid state experiment on proteins. Four-channel multinuclear probes with gradients have been the workhorse in solution NMR for decades, but they have not been available for ssNMR – they have been perceived to be impractically difficult to design and build. The Phase-I demonstrated feasibility of an H/X/Y/Z narrow-bore (NB) MAS probe based on a novel “single-coil” rf circuit optimized for ¹H detection and suitable for use at fields from 7-31 T. The Phase-II probe will be compatible with automated sample exchange, pulsed-field gradients (PFG), NB magnets, and novel NB microwave irradiation methods for Dynamic Nuclear Polarization (DNP). Calculations suggest ²H J-couplings contribute 5-10 Hz to the remaining ¹H line broadening in rigid proteins, and available data suggest the probe-limited resolution (from thermal gradients and magnetism) in commercially available fast-MAS probes has contributed another 6-25 Hz. A 4-channel MAS probe with order- of-magnitude lower thermal gradients that is capable of 2 Hz ¹H resolution on liquids is expected to enable ¹H linewidths below 0.01 ppm on most of the residues in rigid proteins at 900 MHz and above. The novel circuit will also be tunable to virtually all combinations of interest, such as ¹H/¹³C/²H/¹⁵N, ¹H/³¹P/¹³C/²H, ¹H/³¹P/⁷Li/¹³C, ¹H/²⁷Al/²⁹Si/¹⁷O, and ¹H/¹³C/²⁹Si/¹⁰³Rh, thereby making it also invaluable in such areas as metabolism, neurology, materials science, catalysis, and sustainable energy.

HXYZ-g HR-Fast-MAS探针显著改善生物分子结构测定