Awad Wael, Svensson Birkedal Gabriel, Thunnissen Marjolein M G M, Mani Katrin, Logan Derek T
Department of Biochemistry and Structural Biology, Centre for Molecular Protein Science, Lund University, Box 124, 221 00 Lund, Sweden.
Acta Crystallogr D Biol Crystallogr. 2013 Dec;69(Pt 12):2524-33. doi: 10.1107/S0907444913025250. Epub 2013 Nov 19.
The use of controlled dehydration for improvement of protein crystal diffraction quality is increasing in popularity, although there are still relatively few documented examples of success. A study has been carried out to establish whether controlled dehydration could be used to improve the anisotropy of crystals of the core protein of the human proteoglycan glypican-1. Crystals were subjected to controlled dehydration using the HC1 device. The optimal protocol for dehydration was developed by careful investigation of the following parameters: dehydration rate, final relative humidity and total incubation time Tinc. Of these, the most important was shown to be Tinc. After dehydration using the optimal protocol the crystals showed significantly reduced anisotropy and improved electron density, allowing the building of previously disordered parts of the structure.
尽管成功的实例相对较少,但使用可控脱水来提高蛋白质晶体衍射质量的方法正越来越受欢迎。已经开展了一项研究,以确定可控脱水是否可用于改善人蛋白聚糖磷脂酰肌醇蛋白聚糖-1核心蛋白晶体的各向异性。使用HC1设备对晶体进行可控脱水。通过仔细研究以下参数制定了最佳脱水方案:脱水速率、最终相对湿度和总孵育时间Tinc。其中,最重要的是Tinc。使用最佳方案脱水后,晶体的各向异性显著降低,电子密度提高,从而能够构建结构中先前无序的部分。