The effect of bacteriorhodopsin, detergent and hydration on the cubic-to-lamellar phase transition in the monoolein-distearoyl phosphatidyl glycerol-water system.
The cubic phase of monoolein (MO) has successfully been used for crystallization of membrane proteins. It is likely that the transition to a lamellar phase upon dehydration is important for the crystallization process, and that the internal dimensions of the lipid phases (i.e., water pore diameter) are crucial for the inclusion and the diffusion of membrane proteins. In the present study, we investigated the cubic-to-lamellar phase transitions in the MO-water and the MO-distearoyl phosphatidyl glycerol (DSPG) systems. The MO-water system was investigated by means of isothermal sorption and desorption microcalorimetry. We show that the transition from cubic to lamellar phase induced by desorption is driven by entropy. At 25 degrees C, this occurs at a water activity of 0.98 with a transition enthalpy of 860 J/mol (MO). The phase behavior was also investigated in the presence of a small amount of the transmembrane protein bacteriorhodopsin (bR), and a detergent, octyl glucoside (OG), and it was shown that both bR and OG stabilize the lamellar phase. Analogous results were obtained for the MO-DSPG-water system. The latter system resembles the MO-water system in that a cubic-to-lamellar phase transition is induced by dehydration, although the structural properties of these phases are slightly different. Finally, we demonstrate that bR can be crystallized from a cubic phase of MO-DSPG-buffer.
Department of Pharmacy, Uppsala University, Box 580, SE-751 23 Uppsala, Sweden. emma.sparr@fkem1.lu.se
Emma
E
Sparr
Pia
P
Wadsten
Vitaly
V
Kocherbitov
Sven
S
Engström
D001436:Bacteriorhodopsins / Q000494:pharmacology; D002151:Calorimetry; D003902:Detergents / Q000494:pharmacology; D019277:Entropy; D005960:Glucosides / Q000494:pharmacology; D005989:Glycerides / Q000737:chemistry*; D044367:Phase Transition / Q000187:drug effects*; D010715:Phosphatidylglycerols / Q000737:chemistry*; D014867:Water / Q000494:pharmacology
D003902:Detergents; D005960:Glucosides; D005989:Glycerides; D010715:Phosphatidylglycerols; D014867:Water; C018619:octyl-beta-D-glucoside; C084352:distearoyl phosphatidylglycerol; D001436:Bacteriorhodopsins; C005953:monoolein