Cardiolipin, a key component to mimic the E. coli bacterial membrane in model systems revealed by dynamic light scattering and steady-state fluorescence anisotropy
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The phase transition temperatures of several lipidic systems were determined using two different techniques: dynamic light scattering (DLS) and steady-state fluorescence anisotropy, using two fluorescent probes that report different membrane regions (TMA-DPH and DPH). Atomic force microscopy (AFM) was used as a complementary technique to characterize different lipid model systems under study. The systems were chosen due to the increased interest in bacterial membrane studies due to the problem of antibiotic drug resistance. The simpler models studied comprised of mixtures of POPE and POPG lipids, which form a commonly used model system for Escherichia coli membranes. Given the important role of cardiolipin (CL) in natural membranes, a ternary model system, POPE/POPG/CL, was then considered. The results obtained in these mimetic systems were compared with those obtained for the natural systems E. coli polar and total lipid extract. DLS and fluorescence anisotropy are not commonly used to study lipid phase transitions, but it was shown that they can give useful information about the thermotropic behaviors of model systems for bacterial membranes. These two techniques provided very similar results, validating their use as methods to measure phase transitions in lipid model systems. The temperature transitions obtained from these two very different techniques and the AFM results clearly show that cardiolipin is a fundamental component to mimic bacteria membranes. The results suggest that the less commonly used ternary system is a considerably better mimic for natural E. coli membranes than binary lipid mixture.
KeywordsLipid membranes Cardiolipin Dynamic light scattering Fluorescence anisotropy Atomic force microscopy
Dynamic light scattering
Atomic force microscopy
Differential scanning calorimetry
Nuclear magnetic resonance
Fourier transform infrared spectroscopy
N-(2-Hydroxyethyl) piperazine-N′-ethanesulfonic acid
Large unilamellar vesicle
Partial financial support for this work was provided by EU project Translocation MRTN-CT-2005-019335. S. Lopes and C. Neves thank FCT for a SFRH/BPD/34262/2006 and SFRH/BD/61137/2009 fellowship, respectively.
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