Buffer-induced swelling and vesicle budding in binary lipid mixtures of dioleoylphosphatidylcholine:dioleoylphosphatidylethanolamine and dioleoylphosphatidylcholine:lysophosphatidylcholine using small-angle X-ray scattering and ³¹P static NMR.
A large variety of data exists on lipid phase behavior; however, it is mostly in nonbuffered systems over nonbiological temperature ranges. We present biophysical data on lipid mixtures of dioleoylphosphatidylcholine (DOPC), dioleoylphosphatidylethanolamine (DOPE), and lysophosphatidylcholine (LysoPC) examining their behaviors in excess water and buffer systems over the temperature range 4-34 °C. These mixtures are commonly used to investigate the effects of spontaneous curvature on integral membrane proteins. Using small-angle X-ray scattering (SAXS) and (31)P NMR, we observed lamellar and vesicle phases, with the buffer causing an increase in the layer spacing. Increasing amounts of DOPE in a DOPC bilayer decreased the layer spacing of the mesophase, while the opposite trend was observed for increasing amounts of LysoPC. (31)P static NMR was used to analyze the DOPC:LysoPC samples to investigate the vesicle sizes present, with evidence of vesicle budding observed at LysoPC concentrations above 30 mol %. NMR line shapes were fitted using an adapted program accounting for the distortion of the lipids within the magnetic field. The distortion of the vesicle, because of magnetic susceptibility, varied with LysoPC content, and a discontinuity was found in both the water and buffer samples. Generally, the distortion increased with LysoPC content; however, at a ratio of DOPC:LysoPC 60:40, the sample showed a level of distortion of the vesicle similar to that of pure DOPC. This implies an increased flexibility in the membrane at this point. Commonly, the assumption is that for increasing LysoPC concentration there is a reduction in membrane tension, implying that estimations of membrane tension based on spontaneous curvature assumptions may not be accurate.
†Department of Chemistry, Imperial College London, London, SW7 2AZ, United Kingdom.
Hanna M G
HM
Barriga
‡Pfizer Global Research and Development, Sandwich, Kent CT13 9NJ, United Kingdom.
Richard
R
Bazin
†Department of Chemistry, Imperial College London, London, SW7 2AZ, United Kingdom.
Richard H
RH
Templer
†Department of Chemistry, Imperial College London, London, SW7 2AZ, United Kingdom.
Robert V
RV
Law
†Department of Chemistry, Imperial College London, London, SW7 2AZ, United Kingdom.
Oscar
O
Ces
D002021:Buffers; D002462:Cell Membrane / Q000737:chemistry; D008051:Lipid Bilayers / Q000737:chemistry*; D008244:Lysophosphatidylcholines / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy; D010713:Phosphatidylcholines / Q000737:chemistry*; D010714:Phosphatidylethanolamines / Q000737:chemistry*; D053838:Scattering, Small Angle; D014961:X-Ray Diffraction
D002021:Buffers; D008051:Lipid Bilayers; D008244:Lysophosphatidylcholines; D010713:Phosphatidylcholines; D010714:Phosphatidylethanolamines; C094877:dioleoyl phosphatidylethanolamine; C017251:1,2-oleoylphosphatidylcholine