Lipidic cubic phases as matrices for membrane protein crystallization.
This review provides detailed procedures for the crystallization of membrane proteins via the lipidic cubic phase method. Bacteriorhodopsin-specific, hands-on protocols are given for (i) the preparation of bacteriohordopsin from purple membrane by monomerization in octylglucoside and gel filtration chromatography or by selective extraction after pre-treatment with dodecyl-trimethylammonium bromide, (ii) the incorporation of bacteriorhodopsin into lipidic cubic phases by mixing in vials or within coupled syringes and, (iii) the crystallization of bacteriorhodopsin in the lipidic matrix by adding a solid salt or an overlaying with a solution. References for further useful procedures and materials are listed in order to provide biochemists and crystallographers with all information that is necessary to grow crystals of the membrane protein bacteriorhodopsin.
deCODE genetics, BioStructures Group, 7869 NE Day RdW, Bainbridge Island, WA 98110, USA. pnollert@decode.com
Peter
P
Nollert
D001436:Bacteriorhodopsins / Q000737:chemistry*; D002623:Chemistry Techniques, Analytical / Q000379:methods*; D002850:Chromatography, Gel; D003460:Crystallization; D005960:Glucosides; D019617:Halobacterium salinarum / Q000737:chemistry; D008055:Lipids / Q000737:chemistry*; D018717:Purple Membrane / Q000737:chemistry
D005960:Glucosides; D008055:Lipids; C018619:octyl-beta-D-glucoside; D001436:Bacteriorhodopsins