A Novel Approach for Protein Crystallization by a Synthetic Hydrogel with Thermoreversible Gelation Polymer

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• 作者：Shigeru Sugiyama ; Noriko Shimizu ; Gen Sazaki ; Mika Hirose ; Yoshinori Takahashi ; Mihoko Maruyama ; Hiroyoshi Matsumura ; Hiroaki Adachi ; Kazufumi Takano ; Satoshi Murakami ; Tsuyoshi Inoue ; Yusuke Mori
• 刊名：Crystal Growth & Design
• 出版年：2013
• 出版时间：May 1, 2013
• 年：2013
• 卷：13
• 期：5
• 页码：1899-1904
• 全文大小：392K
• 年卷期：v.13,no.5(May 1, 2013)
• ISSN：1528-7505

文摘

Agarose has been utilized in protein crystallization to control nucleation of protein crystals. It reduces convection, prevents crystal sedimentation, and increases tolerance to environmental perturbations, resulting in high-quality protein crystals. However, crystallographers have seldom used agarose hydrogel because it requires preincubating the crystallization solution at high temperatures where a high-temperature-sensitive protein may be inactivated or aggregated. To overcome this disadvantage, we used a thermoreversible gel polymer (TGP) made from synthetic polymer. TGP turns into hydrogel upon warming and liquefies upon cooling. This novel approach enabled us to prepare the crystallization solutions at low temperature (277鈥?83 K) and to crystallize elastase, glucose isomerase, and lysozyme with TGP. We also found that TGP clearly increased the number of elastase, glucose isomerase, and lysozyme crystals. This approach will provide a wide variety of possibilities for protein crystallization in hydrogels.