The lipid composition of Legionella dumoffii membrane modulates the interaction with Galleria mellonella apolipophorin III.
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. (31)P 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.
Department of Genetics and Microbiology, Institute of Microbiology and Biotechnology, Maria Curie-Sklodowska University, Akademicka St. 19, 20-033 Lublin, Poland. Electronic address: marta.szysz@poczta.umcs.lublin.pl.
Marta
M
Palusińska-Szysz
Department of Immunobiology, Institute of Biology and Biochemistry, Maria Curie-Sklodowska University, Akademicka St. 19, 20-033 Lublin, Poland. Electronic address: barabas@poczta.umcs.lublin.pl.
Agnieszka
A
Zdybicka-Barabas
Department of Biophysics, Institute of Physics, Maria Curie-Sklodowska University, Maria Curie-Sklodowska Square 1, 20-031 Lublin, Poland; Department of Biophysics, Institute of Biology and Biochemistry, Maria Curie-Sklodowska University, Akademicka St. 19, 20-033 Lublin, Poland. Electronic address: e.reszczynska@gmail.com.
Emilia
E
Reszczyńska
Department of Biophysics, Institute of Physics, Maria Curie-Sklodowska University, Maria Curie-Sklodowska Square 1, 20-031 Lublin, Poland. Electronic address: rafal.luchowski@umcs.pl.
Rafał
R
Luchowski
Mass Spectrometry Group, Institute of Organic Chemistry Polish Academy of Sciences, Kasprzaka 44/52 St., 01-224 Warsaw, Poland. Electronic address: magdalena.kania@icho.edu.pl.
Magdalena
M
Kania
Division of Bioanalytical Chemistry, Research Center Borstel, Leibniz-Center for Medicine and Biosciences, Parkallee 1-40, 23845 Borstel, Germany. Electronic address: ngisch@fz-borstel.de.
Nicolas
N
Gisch
Division of Bioanalytical Chemistry, Research Center Borstel, Leibniz-Center for Medicine and Biosciences, Parkallee 1-40, 23845 Borstel, Germany. Electronic address: fwaldow@fz-borstel.de.
Franziska
F
Waldow
Department of Analytical Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Gronostajowa 7 St., 30-387 Krakow; Malopolska Centre of Biotechnology, Gronostajowa 7A St., 30-387 Krakow, Poland. Electronic address: pawel.mak@uj.edu.pl.
Paweł
P
Mak
Mass Spectrometry Group, Institute of Organic Chemistry Polish Academy of Sciences, Kasprzaka 44/52 St., 01-224 Warsaw, Poland. Electronic address: witold.danikiewicz@icho.edu.pl.
Witold
W
Danikiewicz
Department of Biophysics, Institute of Physics, Maria Curie-Sklodowska University, Maria Curie-Sklodowska Square 1, 20-031 Lublin, Poland. Electronic address: wieslaw.gruszecki@umcs.pl.
Wiesław I
WI
Gruszecki
Department of Immunobiology, Institute of Biology and Biochemistry, Maria Curie-Sklodowska University, Akademicka St. 19, 20-033 Lublin, Poland. Electronic address: cytryna@poczta.umcs.lublin.pl.
Małgorzata
M
Cytryńska
D000818:Animals; D001053:Apolipoproteins / Q000737:chemistry* / Q000302:isolation & purification; D002462:Cell Membrane / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism; D002794:Choline / Q000494:pharmacology; D005456:Fluorescent Dyes / Q000737:chemistry; D054884:Host-Pathogen Interactions; D006801:Humans; D019476:Insect Proteins / Q000737:chemistry* / Q000302:isolation & purification; D007875:Legionella / Q000737:chemistry* / Q000187:drug effects / Q000254:growth & development / Q000378:metabolism; D008051:Lipid Bilayers / Q000737:chemistry / Q000378:metabolism; D008081:Liposomes / Q000737:chemistry / Q000378:metabolism; D009036:Moths / Q000737:chemistry / Q000382:microbiology*; D010713:Phosphatidylcholines / Q000737:chemistry; D010714:Phosphatidylethanolamines / Q000737:chemistry; D011485:Protein Binding; D013050:Spectrometry, Fluorescence; D017550:Spectroscopy, Fourier Transform Infrared
D001053:Apolipoproteins; D005456:Fluorescent Dyes; D019476:Insect Proteins; D008051:Lipid Bilayers; D008081:Liposomes; D010713:Phosphatidylcholines; D010714:Phosphatidylethanolamines; C042470:apolipophorin III; C483858:phosphatidylethanolamine; D002794:Choline
Apolipophorin III; Fluorescence lifetime imaging microscopy; Fourier transform infrared spectroscopy; Membrane phospholipids; Phosphatidylcholine