Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
8301688 | Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids | 2016 | 13 Pages |
Abstract
Apolipophorin III (apoLp-III), an insect homologue of human apolipoprotein E (apoE), is a widely used model protein in studies on protein-lipid interactions, and anti-Legionella activity of Galleria mellonella apoLp-III has been documented. Interestingly, exogenous choline-cultured Legionella dumoffii cells are considerably more susceptible to apoLp-III than non-supplemented bacteria. In order to explain these differences, we performed, for the first time, a detailed analysis of L. dumoffii lipids and a comparative lipidomic analysis of membranes of bacteria grown without and in the presence of exogenous choline. 31P NMR analysis of L. dumoffii phospholipids (PLs) revealed a considerable increase in the phosphatidylcholine (PC) content in bacteria cultured on choline medium and a decrease in the phosphatidylethanolamine (PE) content in approximately the same range. The interactions of G. mellonella apoLp-III with lipid bilayer membranes prepared from PLs extracted from non- and choline-supplemented L. dumoffii cells were examined in detail by means of attenuated total reflection- and linear dichroism-Fourier transform infrared spectroscopy. Furthermore, the kinetics of apoLp-III binding to liposomes formed from L. dumoffii PLs was analysed by fluorescence correlation spectroscopy and fluorescence lifetime imaging microscopy using fluorescently labelled G. mellonella apoLp-III. Our results indicated enhanced binding of apoLp-III to and deeper penetration into lipid membranes formed from PLs extracted from the choline-supplemented bacteria, i.e. characterized by an increased PC/PE ratio. This could explain, at least in part, the higher susceptibility of choline-cultured L. dumoffii to G. mellonella apoLp-III.
Keywords
dMPCgas liquid chromatography/mass spectrometryGLC/MSMMPEdimyristoylphosphatidylcholineApolipophorin IIIDPPCTLCFITCnuclear magnetic resonanceFLIMLC/ESI-MSNMRdipalmitoylphosphatidylcholineFTIRFourier transform infrared spectroscopyMembrane phospholipidsphosphatidylcholinephosphatidylglycerolphosphatidylethanolaminefluorescein isothiocyanatefluorescence lifetime imaging microscopyCardiolipinthin-layer chromatography
Related Topics
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Biochemistry, Genetics and Molecular Biology
Biochemistry
Authors
Marta PalusiÅska-Szysz, Agnieszka Zdybicka-Barabas, Emilia ReszczyÅska, RafaÅ Luchowski, Magdalena Kania, Nicolas Gisch, Franziska Waldow, PaweÅ Mak, Witold Danikiewicz, WiesÅaw I. Gruszecki, MaÅgorzata CytryÅska,