A Single Entity Method for Controlled Nucleation and Crystal Growth

控制成核和晶体生长的单一实体方法

• 批准号: 10720470
• 负责人: Gangli Wang
• 金额: $ 36.45万
• 依托单位: GEORGIA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2027-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10720470
• 关键词: 3-Dimensional; Adopted; Area; Binding; Biological Process; Biomedical Research; Cell Nucleus; Chemicals; Complex; Crystal Formation; Crystallization; Data Collection; Development; Diffusion; Disadvantaged; Disease; Drug Design; Electron Microscopy; Embryo; Feedback; Formulation; Future; Goals; Growth; Habits; Hydrogen; Individual; Inherited; Insulin; Ions; Kinetics; Lipids; Liquid substance; Location; Magic; Measurement; Membrane Proteins; Methodology; Methods; Monitor; Muramidase; Names; Nature; Needles; Neutrons; Noise; Nucleic Acids; Optical Methods; Optics; Pharmacologic Substance; Phase; Phase Transition; Physiological; Play; Preparation; Process; Program Development; Proteins; Protons; Quantitative Evaluations; R-factor; Reproducibility; Research; Resolution; Roentgen Rays; Role; Sampling; Shapes; Signal Transduction; Site; Sodium Chloride; Solvents; Source; Structure; Surface; System; Techniques; Temperature; Thermodynamics; Thinness; Time; Transport Process; Uncertainty; Variant; X-Ray Crystallography; active control; biomacromolecule; design; drug development; improved; insight; macromolecule; migration; nano; nanodevice; nanoscale; outcome prediction; predictive signature; programs; prototype; real time monitoring; success; technology development; tool; two-dimensional; vapor

The structures of biomacromolecules at atomic resolution (< 2.0-2.5 Å) are of enormous importance to understand their physiological functions and roles in diseases. An exemplary critical need of high atomic resolution is to resolve the location of proton/hydrogen which plays vital roles in various biological processes. Deuteration renders neutron scattering techniques unique advantages in high contrast (signal/background) to locate D/H. Like X-ray crystallography which has contributed majority of known biomolecule structures, high quality single crystals are the prerequisites for both X-ray and neutron data collection. It is worth mentioning that despite the recent progresses in electron microscopy techniques, true atomic resolution remains a formidable challenge to achieve. Lower resolution structures are associated with ambiguity and could mislead basic biomedical research as well as drug design/development applications. With the understanding on the fundamental limitations and technical hurdles associated with currently adopted ensemble-based methods, we propose to develop a single-entity method (named NanoAC) which will offer unprecedented capability to synthesize crystals one at a time, under real-time monitoring and with predictive crystal quality. A single nanotip will be employed to spatially confine supersaturation as the sole nucleation site. Electroanalytical and optical methods will monitor the whole crystallization process in real-time to capture quantitative signatures for the nucleation and crystal growth at single entity resolutions. Those signatures will enable active controls in kinetic transitions, and be quantitatively correlated with its diffraction quality and/or crystal habits. The insights will inform crystal synthesis such that nucleation kinetics and growth rates of each individual crystal will be finetune to improve crystal quality and to tune crystal size/habits. Prototype soluble proteins, nucleic acids and membrane proteins will be used as defined in this early-stage technology development program. The new toolbox, once established, will provide paradigm-shift capabilities to improve the crystal quality in diffraction and size/habit controls, to tackle challenging material systems currently not-crystallizable, and also feature high efficiency in time and/or materials. The overarching goal will be pursued through three interrelated aims. Aim 1 will establish real-time monitoring signatures for the generalization of NanoAC to crystallize soluble biomacromolecules and complexes. Aim 2 will correlate diffraction quality and crystal habits with monitoring signatures. Aim 3 will further develop single nanopipettes as ‘magic wand’ to crystallize membrane proteins.

原子分辨率（<2.0-2.5Å）在原子分辨率上的生物乳清分子的结构至关重要 了解他们的身体功能和在疾病中的作用。高原子的示例性临界需求 解决方案是解决在各种生物过程中起着至关重要的作用的质子/氢的位置。 侵蚀使中子散射技术在高对比度（信号/背景）中具有独特的优势 找到D/H。像X射线晶体学，贡献了大多数已知的生物分子结构，高 优质的单晶是X射线和中子数据收集的先决条件。值得一提的 尽管最近的电子显微镜技术取得了进展，但真原子分辨率仍然很强大 挑战要实现。较低的分辨率结构与歧义有关，可能误导基本 生物医学研究以及药物设计/开发应用。有了对 与当前采用的基于合奏的方法相关的基本限制和技术障碍，我们 提出开发单一实体方法（名为nanoac）的建议，该方法将提供前所未有的能力 在实时监测和预测晶体质量下，一次合成一个晶体。单个纳米 将在空间上将其作为唯一的核部位进行空间限制。电分析和光学 方法将实时监视整个结晶过程，以捕获用于的定量特征 单实体分辨率的成核和晶体生长。这些签名将在动力学中实现主动控制 过渡，并与其衍射质量和/或晶体习惯定量相关。见解将告知 晶体合成使每个单个晶体的成核动力学和生长速率将是未来的 提高晶体质量并调整晶体尺寸/习惯。原型固体蛋白，核酸和膜 蛋白质将按照该早期技术开发计划的定义使用。新工具箱一次 建立的，将提供范式移位能力，以提高衍射和尺寸/习惯的晶体质量 控件，以应对目前无法结晶的挑战材料系统，并且还具有很高的效率 时间和/或材料。总体目标将通过三个相互关联的目标实现。 AIM 1将建立 实时监测特征，以概括纳米AC，以使固体生物大分子和 复合物。 AIM 2将将衍射质量和晶体习惯与监测特征相关联。 AIM 3将进一步 开发单个纳米夹作为“魔杖”，以使膜蛋白结晶。