Division of Biotechnology, College of Life Sciences and Biotechnology, Korea University, Seoul 02841, Korea.
Institute of Life Science and Natural Resources, Korea University, Seoul 02841, Korea.
Int J Mol Sci. 2019 Mar 4;20(5):1094. doi: 10.3390/ijms20051094.
X-ray crystallographic methods can be used to visualize macromolecules at high resolution. This provides an understanding of molecular mechanisms and an insight into drug development and rational engineering of enzymes used in the industry. Although conventional synchrotron-based X-ray crystallography remains a powerful tool for understanding molecular function, it has experimental limitations, including radiation damage, cryogenic temperature, and static structural information. Serial femtosecond crystallography (SFX) using X-ray free electron laser (XFEL) and serial millisecond crystallography (SMX) using synchrotron X-ray have recently gained attention as research methods for visualizing macromolecules at room temperature without causing or reducing radiation damage, respectively. These techniques provide more biologically relevant structures than traditional X-ray crystallography at cryogenic temperatures using a single crystal. Serial femtosecond crystallography techniques visualize the dynamics of macromolecules through time-resolved experiments. In serial crystallography (SX), one of the most important aspects is the delivery of crystal samples efficiently, reliably, and continuously to an X-ray interaction point. A viscous delivery medium, such as a carrier matrix, dramatically reduces sample consumption, contributing to the success of SX experiments. This review discusses the preparation and criteria for the selection and development of a sample delivery medium and its application for SX.
X 射线晶体学方法可用于高分辨率可视化大分子。这提供了对分子机制的理解，并深入了解药物开发和工业中使用的酶的合理工程。虽然基于传统同步加速器的 X 射线晶体学仍然是理解分子功能的强大工具，但它具有实验限制，包括辐射损伤、低温和静态结构信息。使用 X 射线自由电子激光 (XFEL) 的连续飞秒晶体学 (SFX) 和使用同步加速器 X 射线的连续毫秒晶体学 (SMX) 最近作为在室温下可视化大分子而不引起或减少辐射损伤的研究方法受到关注。与使用单晶的低温传统 X 射线晶体学相比，这些技术提供了更具生物学相关性的结构。连续飞秒晶体学技术通过时间分辨实验可视化大分子的动力学。在连续晶体学 (SX) 中，最重要的方面之一是将晶体样品有效地、可靠地和连续地输送到 X 射线相互作用点。粘性输送介质，如载体基质，大大减少了样品消耗，有助于 SX 实验的成功。本综述讨论了样品输送介质的制备和选择标准及其在 SX 中的应用。
