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Protein Crystallization Using Room Temperature Ionic FluidsThe ionic liquids (ILs) 1-butyl-3-methylimidizolium chloride (C4mim-C1), 1-butyl-3- methylimidizolium diethyleneglycol monomethylethersulfate ([C4mim]DEMGS), and 1-butyl-1 -methylpyrollidinium dihydrogenphosphate ([p1,4]dhp) were tested for their effects on the crystallization of the proteins canavalin, beta-lactoglobulin B, xylanase, and glucose isomerase, using a standard high throughput screen. The crystallization experiments were set up with the ILs added to the protein solutions at 0.2 and 0.4 M final concentrations. Crystallization droplets were set up at three proteixprecipitant ratios (1:1, 2:1, and 4:l), which served to progressively dilute the effects of the screen components while increasing the equilibrium protein and IL concentrations. Crystals were obtained for all four proteins at a number of conditions where they were not obtained from the IL-free control experiment. Over half of the protein-IL combinations tested had more successful outcomes than negative, where the IL-free crystallization was better than the corresponding IL-containing outcome, relative to the control. One of the most common causes of a negative outcome was solubilization of the protein by the IL, resulting in a clear drop. In one instance, we were able to use the IL-induced solubilizing to obtain beta-lactoglobulin B crystals from conditions that gave precipitated protein in the absence of IL. The results suggest that it may be feasible to develop ILs specifically for the task of macromolecule crystallization.
Document ID
20070002061
Acquisition Source
Marshall Space Flight Center
Document Type
Preprint (Draft being sent to journal)
Authors
Pusey, Marc L.
(NASA Marshall Space Flight Center Huntsville, AL, United States)
Paley, Mark Steve
(AZ Technology, Inc. Huntsville, AL, United States)
Turner, Megan B.
(Alabama Univ. Tuscaloosa, AL, United States)
Rogers, Robin D.
(Alabama Univ. Tuscaloosa, AL, United States)
Date Acquired
August 24, 2013
Publication Date
January 1, 2006
Subject Category
Inorganic, Organic And Physical Chemistry
Funding Number(s)
CONTRACT_GRANT: GM071581-01
Distribution Limits
Public
Copyright
Work of the US Gov. Public Use Permitted.

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