Novel Device and Strategy for Growing Large, High-Quality Protein Crystals by Controlling Crystallization Conditions

结晶 蛋白质结晶 材料科学 Crystal(编程语言) 聚乙二醇 晶体生长 结晶水 化学工程 结晶学 衍射 同步加速器 PEG比率 中子衍射 化学 晶体结构 光学 无机化学 计算机科学 物理 工程类 冶金 程序设计语言
作者
Naoki Tanigawa,Sachiko Takahashi,Bangbo Yan,Masayuki Kamo,Naoki Furubayashi,Koji Kubota,Koji Inaka,Hiroaki Tanaka
出处
期刊:Crystals [Multidisciplinary Digital Publishing Institute]
卷期号:11 (11): 1311-1311 被引量:1
标识
DOI:10.3390/cryst11111311
摘要

Neutron diffraction experiments are informative for determining the locations of hydrogen atoms in protein molecules; however, much larger crystals are needed than those required for X-ray diffraction. Thus, additional techniques are required to grow larger crystals. Here, a unique crystallization device and strategy for growing large protein crystals are introduced. The device uses two micropumps to control crystal growth by altering the precipitant concentration and regulating the pinpoint injection of dry air flow to the crystallization cell. Furthermore, the crystal growth can be observed in real time. Preliminary microbatch crystallization experiments at various concentration ranges of polyethylene glycol (PEG) 4000 and sodium chloride were first performed to elucidate optimized crystallization conditions. Based on these results, a device to precisely control the sodium chloride and PEG concentrations and the supply of dry air to the crystallization cell was used, and 1.8 mm lysozyme and 1.5 mm alpha-amylase crystals with good reproducibility were obtained. X-ray data sets of both crystals were collected at room temperature at BL2S1 of the Aichi Synchrotron Radiation Center and confirmed that these crystals were of high quality. Therefore, this crystallization device and strategy were effective for growing large, high-quality protein crystals.
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