Journal
JOURNAL OF APPLIED CRYSTALLOGRAPHY
Volume 45, Issue -, Pages 1330-1333Publisher
INT UNION CRYSTALLOGRAPHY
DOI: 10.1107/S0021889812037880
Keywords
crystallization; in meso crystallogenesis; lipids; membrane proteins; mesophase; receptors
Categories
Funding
- Science Foundation Ireland [07/IN.1/B1836]
- National Institutes of Health [GM75915, P50GM073210, U54GM094599]
- FP7 COST Action [CM0902]
- Science Foundation Ireland (SFI) [07/IN.1/B1836] Funding Source: Science Foundation Ireland (SFI)
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A simple and inexpensive protocol for producing crystals in the sticky and viscous mesophase used for membrane protein crystallization by the in meso method is described. It provides crystals that appear within 15-30 min of setup at 293 K. The protocol gives the experimenter a convenient way of gaining familiarity and a level of comfort with the lipidic cubic mesophase, which can be daunting as a material when first encountered. Having used the protocol to produce crystals of the test protein, lysozyme, the experimenter can proceed with confidence to apply the method to more valuable membrane (and soluble) protein targets. The glass sandwich plates prepared using this robust protocol can further be used to practice harvesting and snap-cooling of in meso-grown crystals, to explore diffraction data collection with mesophase-embedded crystals, and for an assortment of quality control and calibration applications when used in combination with a crystallization robot.
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