Department of Molecular Biology and Biochemistry, University of CA Irvine, Irvine, CA, United States; The Scripps Research Institute Florida, Jupiter, FL, United States.
Methods Enzymol. 2024;709:1-27. doi: 10.1016/bs.mie.2024.10.003. Epub 2024 Oct 29.
The production of enzyme microcrystals for time resolved serial crystallography employing free electron laser or synchrotron radiation is a relatively new variation on traditional macromolecular crystallization for conventional single crystal X-ray analysis. While the fundamentals of macromolecular crystal growth are the same, some modifications and special considerations are in order if the objective is to produce uniform size, microcrystals in very large numbers for serial data collection. Presented here are the basic principles of protein crystal growth with particular attention to the approaches best employed to achieve the goal of microcrystals and some novel techniques, as well as old, that may be useful. Also discussed are the advantages of particular precipitants and certain methods of growing protein crystals that might be advantageous for serial data recording.
利用自由电子激光或同步辐射生产酶微晶体进行时分辨连续晶体学是传统大分子晶体学用于常规单晶 X 射线分析的相对较新的变体。虽然大分子晶体生长的基本原理是相同的,但是如果目标是生产大量均匀大小的微晶体进行连续数据收集,则需要进行一些修改和特殊考虑。本文介绍了蛋白质晶体生长的基本原理,特别关注了实现微晶体目标的最佳方法以及一些新的和旧的可能有用的技术。还讨论了特定沉淀剂的优势以及可能有利于连续数据记录的某些蛋白质晶体生长方法。