Abstract
In this study, we explore the use of line FRAP to detect diffusion in synthetic lipid membranes. The study of the dynamics of these membrane lipids can, however, be challenging. The diffusion in two different synthetic membranes consisting of the lipid mixtures 1:1 DOPC:DPPC and 2:2:1 DOPC:DPPC:Cholesterol was studied with line FRAP. A correlation between diffusion coefficient and temperature was found to be dependent on the morphology of the membrane. We suggest line FRAP as a promising accessible and simple technique to study diffusion in plasma membranes.
Highlights
The descriptions of the fluid-mosaic model [1,2] led to an increased focus on understanding the lipid dynamics in biological membranes and gave rise to multiple ways of investigating these;a very popular technique is the use of synthetic membranes
The first diffusion coefficient measured from all line Fluorescence recovery after photobleaching (FRAP) data was 5 to 10-fold higher than in comparable synthetic membranes, reaching from 16 to 45 μm2 /s [23]; this is expected to be due to free fluorophores in the buffer solution surrounding the membrane and will be neglected in the further data analysis
The second diffusion coefficient shows a correlation to the temperature in the synthetic membrane containing 2:2:1 DOPC:DPPC:Cholesterol
Summary
The descriptions of the fluid-mosaic model [1,2] led to an increased focus on understanding the lipid dynamics in biological membranes and gave rise to multiple ways of investigating these;a very popular technique is the use of synthetic membranes. When designing a synthetic membrane system, it is very important to use the relevant equivalence diagrams, with the most effective being the ternary equivalence diagrams. It should be taken into consideration that these diagrams are temperature dependent Often it is only a small region of the equivalence diagram that results in the desired membrane properties. A common lipid mixture when working with synthetic membranes is a mix of DOPC (1,2-dioleoyl-sn-glycero-3-phosphocholine), DPPC (1,2-dipalmitoyl-sn-glycero-3-phosphocholine) and cholesterol. This mixture is ideal as a synthetic membrane system as it has a relatively large area in the equivalence diagram that fulfills the desired similarities to real plasma membranes [8,9].
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