蛋白质结晶条件的筛选策略研究进展

Research Progresses in Screening Strategies of Protein Crystallization

蛋白质分子的三维结构是生命科学研究中极为重要的信息,X射线单晶衍射技术是目前获得结构信息最主要的手段,但如何筛选到第一个蛋白晶体是该技术必需的第一步,也是制约结构生物学发展的主要瓶颈问题之一。现在一般通过规模筛选的方法从众多的溶液中筛选出可结晶的条件,但是工作量较大,效率也不高。回顾了近年来在提高结晶筛选效率方向取得的成就,主要表现在2个方面:一是在传统结晶方法和试剂基础上发展起来的一系列新的高效的结晶技术和筛选试剂盒;二是从生物学、物理学和化学等角度提出的一些提高结晶筛选效率的新技术,主要包括通过分子工程改造蛋白、提高蛋白溶液的稳定和均一性、导入籽晶、共结晶、改善结晶界面和变温筛选等技术。最后展望了该领域未来的发展趋势。

The structural information of protein molecules is very important to life science, mainly determined by X-ray crystallography for the studies of their functions, but obtaining the first protein crystals is one crucial step, which is often referred to as a ＂bottleneck＂ for structure determination of protein molecules using X-ray diffraction technique. Usually, the first step to obtain diffractable crystals is to get crystallization conditions by screening from hundreds or even thousands of reagents, but it is time and labor cost. Recently, great progresses have been made in this field. In this paper, the progresses in the screening strategies of protein crystallization are reviewed in two aspects： Firstly, some new and efficient screening methods and kits are improved based on the conventional ones; secondly, many new technologies and methods in biology, physics and chemical are developed and the success rate of crystallization screening is enhanced remarkably, including the reconstructing protein by molecular engineering, improving the stability and homogeneity of protein solution, seeding, cocrystallization, improving the interface of crystallization and cycling temperature strategy, and also the future expectancy is discussed.