Low amounts of sucrose are sufficient to depress the phase transition temperature of dry phosphatidylcholine, but not for lyoprotection of liposomes.
Disaccharides such as sucrose and trehalose play an important role in stabilizing cellular structures during dehydration. In fact, most organisms that are able to survive desiccation accumulate high concentrations of sugars in their cells. The mechanisms involved in the stabilization of cellular membranes in the dry state have been investigated using model membranes, such as phosphatidylcholine liposomes. It has been proposed that the lyoprotection of liposomes depends on the depression of the gel to liquid-crystalline phase transition temperature (T(m)) of the dry membranes below ambient and on the prevention of membrane fusion by sugar glass formation, because both lead to leakage of soluble content from the liposomes. Since fusion is prevented at lower sugar/lipid mass ratios than leakage, it has been assumed that more sugar is needed to depress T(m) than to prevent fusion. Here, we show that this is not the case. In air-dried egg phosphatidylcholine liposomes, T(m) is depressed by >60 degrees C at sucrose/lipid mass ratios 10-fold lower than those needed to depress fusion to below 20%. In fact, T(m) is significantly reduced at mass ratios where no bulk sugar glass phase is detectable by Fourier transform infrared spectroscopy or differential scanning calorimetry. A detailed analysis of the interactions of sucrose with the P=O, C=O, and choline groups of the lipid and a comparison to published data on water binding to phospholipids suggests that T(m) is reduced by sucrose through a "water replacement" mechanism. However, the sucrose/lipid mass ratios necessary to prevent leakage exceed those necessary to prevent both phase transitions and membrane fusion. We hypothesize that kinetic phenomena during dehydration and rehydration may be responsible for this discrepancy.
Cacela
Constança
C
Max-Planck-Institut für Molekulare Pflanzenphysiologie, D-14424 Potsdam, Germany.
Hincha
Dirk K
DK
D002152:Calorimetry, Differential Scanning; D002794:Choline / Q000737:chemistry; D003681:Dehydration; D005782:Gels; D006860:Hydrogen Bonding; D008081:Liposomes / Q000737:chemistry*; D008561:Membrane Fusion; D044367:Phase Transition; D010713:Phosphatidylcholines / Q000737:chemistry*; D017550:Spectroscopy, Fourier Transform Infrared; D013395:Sucrose / Q000737:chemistry*; D013816:Thermodynamics; D044366:Transition Temperature; D014867:Water / Q000737:chemistry
D005782:Gels; D008081:Liposomes; D010713:Phosphatidylcholines; D014867:Water; D013395:Sucrose; D002794:Choline