Extensive bilayer perforation coupled with the phase transition region of an anionic phospholipid.
At low ionic strength dimyristoylphosphatidylglycerol (DMPG) exhibits a broad phase transition region characterized by several superimposed calorimetric peaks. Peculiar properties, such as sample transparency, are observed only in the transition region. In this work we use differential scanning calorimetry (DSC), turbidity, and optical microscopy to study the narrowing of the transition region with the increase of ionic strength (0-500 mM NaCl). Upon addition of salt, the temperature extension of the transition region is reduced, and the number of calorimetric peaks decreases until a single cooperative event at T(m) = 23 degrees C is observed in the presence of 500 mM NaCl. The transition region is always coupled with a decrease in turbidity, but a transparent region is detected within the melting process only in the presence of up to 20 mM NaCl. The vanishing of the transparent region is associated with one of the calorimetric peaks. Optical microscopy of giant vesicles shows that bilayers first rupture when the transition region is reached and subsequently lose optical contrast. Fluorescence microscopy reveals a blurry and undefined image in the transparent region, suggesting a different lipid self-assembly. Overall sample turbidity can be directly related to the bilayer optical contrast. Our observations are discussed in terms of the bilayer being perforated along the transition region. In the narrower temperature interval of the transparent region, dependent on the ionic strength, the perforation is extensive and the bilayer completely loses the optical contrast.
Riske
Karin A
KA
Departamento de Biofisica, Universidade Federal de São Paulo, CEP 04023-062, São Paulo, SP, Brazil. kar@biofis.epm.br
Amaral
Lia Q
LQ
Lamy
M Teresa
MT
D000838:Anions; D002151:Calorimetry / Q000379:methods; D002152:Calorimetry, Differential Scanning / Q000379:methods; D002462:Cell Membrane / Q000378:metabolism; D007477:Ions; D008051:Lipid Bilayers / Q000737:chemistry*; D008055:Lipids / Q000737:chemistry; D008853:Microscopy / Q000379:methods; D008856:Microscopy, Fluorescence / Q000379:methods; D009391:Nephelometry and Turbidimetry / Q000379:methods; D044367:Phase Transition; D010715:Phosphatidylglycerols / Q000737:chemistry; D010743:Phospholipids / Q000737:chemistry*; D012965:Sodium Chloride / Q000737:chemistry; D013696:Temperature
D000838:Anions; D007477:Ions; D008051:Lipid Bilayers; D008055:Lipids; D010715:Phosphatidylglycerols; D010743:Phospholipids; D012965:Sodium Chloride; C002773:dimyristoylphosphatidylglycerol