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Biophys J, July 2001, p. 225-242, Vol. 81, No. 1
Biochemistry, Biophysics, and Chemistry, The Ohio State University, Columbus, Ohio 43210 USA
The in meso method for growing crystals of
membrane proteins uses a spontaneously forming lipidic cubic mesophase.
The detergent-solubilized protein is dispersed with lipid, typically
monoolein, and in so doing the cubic phase self-assembles. A
precipitant is added to trigger crystal nucleation and growth. The
commercial screen solution series are convenient for use in
crystallization trials. The aim of this study was to determine which of
the Hampton Screen and Screen 2 series of solutions are compatible with
the in meso method. These screens contain components any
of which could destroy the cubic phase. X-ray diffraction was used for
phase identification and for microstructure characterization. The study
was done at 4°C and at 20°C. Two types of sample preparations were
examined. One used an excess of half-strength screen solution (Prep.
1). The other used a limiting quantity of undiluted screen solution (Prep. 2). At 20°C, over 90% of the screen solutions produced the
cubic phase with Prep. 1. This figure dropped to 50% with Prep. 2. In
contrast, 50 to 60% of the screens were cubic phase compatible at
4°C under Prep. 1 conditions. The figure fell to 25% with Prep. 2. The mode of action of the diverse screen components are explained on
the basis of the phase properties of the monoolein/water system.
Biophys J, July 2001, p. 225-242, Vol. 81, No. 1
© 2001 by the Biophysical Society 0006-3495/01/07/225/18 $2.00
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