Protein Crystallization Using Room Temperature Ionic Liquids
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- 作者：Marc L. Pusey ; Mark Steve Paley ; Megan B. Turner ; Robin D. Rogers
- 刊名：Crystal Growth & Design
- 出版年：2007
- 出版时间：April 2007
- 年：2007
- 卷：7
- 期：4
- 页码：787 - 793
- 全文大小：949K
- 年卷期：v.7,no.4(April 2007)
- ISSN：1528-7505
文摘
The ionic liquids (ILs) 1-butyl-3-methylimidizolium chloride ([C4mim]Cl), 1-butyl-3-methylimidizolium 2(2-methoxyethoxy)ethylsulfate ([C4mim][MDEGSO4]), and 1-butyl-1-methylpyrollidinium dihydrogenphosphate ([p1,4][DHP]) weretested for their effects on the crystallization of the proteins canavalin, 
-lactoglobulin B, xylanase, and glucose isomerase, using astandard high throughput screen. The crystallization experiments were set up with the ILs added to the protein solutions at 0.2 and0.4 M final concentrations. Crystallization droplets were set up at three protein/precipitant ratios (1:1, 2:1, and 4:1), which servedto progressively dilute the effects of the screen components while increasing the equilibrium protein and IL concentrations. Crystalswere obtained for all four proteins at a number of conditions where they were not obtained from IL-free control experiments. Overhalf of the protein-IL combinations tested had more successful outcomes than negative outcomes, where the IL-free crystallizationwas better than the corresponding IL-containing outcome, relative to the control. One of the most common causes of a negativeoutcome was solubilization of the protein by the IL, resulting in a clear drop. In one instance, we were able to use the IL-inducedsolubilizing to obtain -lactoglobulin B crystals from conditions that gave precipitated protein in the absence of IL. The resultssuggest that it may be feasible to develop ILs specifically for the task of macromolecule crystallization.
-lactoglobulin B, xylanase, and glucose isomerase, using astandard high throughput screen. The crystallization experiments were set up with the ILs added to the protein solutions at 0.2 and0.4 M final concentrations. Crystallization droplets were set up at three protein/precipitant ratios (1:1, 2:1, and 4:1), which servedto progressively dilute the effects of the screen components while increasing the equilibrium protein and IL concentrations. Crystalswere obtained for all four proteins at a number of conditions where they were not obtained from IL-free control experiments. Overhalf of the protein-IL combinations tested had more successful outcomes than negative outcomes, where the IL-free crystallizationwas better than the corresponding IL-containing outcome, relative to the control. One of the most common causes of a negativeoutcome was solubilization of the protein by the IL, resulting in a clear drop. In one instance, we were able to use the IL-inducedsolubilizing to obtain -lactoglobulin B crystals from conditions that gave precipitated protein in the absence of IL. The resultssuggest that it may be feasible to develop ILs specifically for the task of macromolecule crystallization.