Key Laboratory for Space Bioscience and Biotechnology, School of Life Sciences, Northwestern Polytechnical University, Xi'an, 710072, China.
Appl Microbiol Biotechnol. 2021 Apr;105(7):2759-2773. doi: 10.1007/s00253-021-11188-z. Epub 2021 Mar 8.
The self-assembly of biomacromolecules is an extremely important process. It is potentially useful in the fields of life science and materials science. To carry out the study on the self-assembly of proteins, it is necessary to find out the suitable self-assembly conditions, which have always been a challenging task in practice. Inspired by the screening technique in the field of protein crystallization, we proposed using the same screening technique for seeking suitable protein self-assembly conditions. Based on this consideration, we selected 5 proteins (β-lactoglobulin, hemoglobin, pepsin, lysozyme, α-chymotrypsinogen (II) A) together with 5 screening kits (Index, BML, Morpheus, JCSG, PEG/Ion Screen) to investigate the performance of these crystallization screening techniques in order to discover new optimized conditions of protein self-assembly. The screens were all kept at 293 K for certain days, and were analyzed using optical microscope, scanning electron microscope, transmission electron microscope, atomic force microscope, fluorescence microscope, and atomic absorption spectroscope. The results demonstrated that the method of protein crystallization screening can be successfully applied in the screening of self-assembly conditions. This method is fast, high throughput, and easily implemented in an automated system, with a low protein consumption feature. These results suggested that such strategy can be applied to finding new conditions or forms in routine research of protein self-assembly. KEY POINTS: • Protein crystallization screening method is successfully applied in the screening of self-assembly conditions. • This screening method can be applied on various kinds of proteins and possess a feature of low protein consumption. • This screening method is fast, high throughput, and easily implemented in an automated system.
生物大分子的自组装是一个极其重要的过程。它在生命科学和材料科学领域具有潜在的应用价值。为了开展蛋白质自组装的研究，有必要找出合适的自组装条件，这在实践中一直是一项具有挑战性的任务。受蛋白质结晶筛选技术的启发，我们提出使用相同的筛选技术来寻找合适的蛋白质自组装条件。基于这一考虑，我们选择了 5 种蛋白质（β-乳球蛋白、血红蛋白、胃蛋白酶、溶菌酶、α-糜蛋白酶原（II）A）和 5 种筛选试剂盒（Index、BML、Morpheus、JCSG、PEG/Ion Screen），以研究这些结晶筛选技术的性能，从而发现蛋白质自组装的新优化条件。所有筛选均在 293 K 下保持一定天数，并使用光学显微镜、扫描电子显微镜、透射电子显微镜、原子力显微镜、荧光显微镜和原子吸收光谱仪进行分析。结果表明，蛋白质结晶筛选方法可成功应用于自组装条件的筛选。该方法快速、高通量，易于在自动化系统中实现，且具有低蛋白消耗的特点。这些结果表明，这种策略可应用于常规蛋白质自组装研究中寻找新的条件或形式。 关键点： • 蛋白质结晶筛选方法成功应用于自组装条件的筛选。 • 该筛选方法适用于各种蛋白质，具有低蛋白消耗的特点。 • 该筛选方法快速、高通量，易于在自动化系统中实现。
