Moreno Abel
Instituto de Química, Universidad Nacional Autónoma de Mexico, Av. Universidad 3000, Cd.Mx., Mexico City, 04510, Mexico.
Methods Mol Biol. 2017;1607:51-76. doi: 10.1007/978-1-4939-7000-1_3.
This chapter provides a review of different advanced methods that help to increase the success rate of a crystallization project, by producing larger and higher quality single crystals for determination of macromolecular structures by crystallographic methods. For this purpose, the chapter is divided into three parts. The first part deals with the fundamentals for understanding the crystallization process through different strategies based on physical and chemical approaches. The second part presents new approaches involved in more sophisticated methods not only for growing protein crystals but also for controlling the size and orientation of crystals through utilization of electromagnetic fields and other advanced techniques. The last section deals with three different aspects: the importance of microgravity, the use of ligands to stabilize proteins, and the use of microfluidics to obtain protein crystals. All these advanced methods will allow the readers to obtain suitable crystalline samples for high-resolution X-ray and neutron crystallography.
本章综述了不同的先进方法,这些方法通过制备更大、质量更高的单晶,以用于通过晶体学方法测定大分子结构,从而有助于提高结晶项目的成功率。为此,本章分为三个部分。第一部分通过基于物理和化学方法的不同策略,阐述了理解结晶过程的基本原理。第二部分介绍了更复杂方法中涉及的新方法,这些方法不仅用于生长蛋白质晶体,还通过利用电磁场和其他先进技术来控制晶体的大小和取向。最后一部分涉及三个不同方面:微重力的重要性、使用配体稳定蛋白质以及使用微流体技术获得蛋白质晶体。所有这些先进方法将使读者能够获得适用于高分辨率X射线和中子晶体学的晶体样品。