Receptor-independent interaction of bacterial lipopolysaccharide with lipid and lymphocyte membranes; the role of cholesterol.
Lipopolysaccharide (LPS) is a major constituent of bacterial outer membranes where it makes up the bulk of the outer leaflet and plays a key role as determinant of bacterial interactions with the host. Membrane-free LPS is known to activate T-lymphocytes through interactions with Toll-like receptor 4 via multiprotein complexes. In the present study, we investigate the role of cholesterol and membrane heterogeneities as facilitators of receptor-independent LPS binding and insertion, which underpin bacterial interactions with the host in symbiosis, pathogenesis and cell invasion. We use fluorescence spectroscopy to investigate the interactions of membrane-free LPS from intestinal gram-negative organisms with cholesterol-containing model membranes and with T-lymphocytes. LPS preparations from Klebsiella pneumoniae and Salmonella enterica were found to bind preferentially to mixed lipid membranes by comparison to pure PC bilayers. The same was observed for LPS from the symbiote Escherichia coli but with an order of magnitude higher dissociation constant. Insertion of LPS into model membranes confirmed the preference for sphingomyelin/cholesterol-containing systems. LPS insertion into Jurkat T-lymphocyte membranes reveals that they have a significantly greater LPS-binding capacity by comparison to methyl-β-cyclodextrin cholesterol-depleted lymphocyte membranes, albeit at slightly lower binding rates.
School of Biomedical Sciences, University of Nottingham, Nottingham, United Kingdom.
Filip
F
Ciesielski
Benjamin
B
Davis
Michael
M
Rittig
Boyan B
BB
Bonev
Paul
P
O'Shea
D001667:Binding, Competitive; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism*; D002784:Cholesterol / Q000737:chemistry / Q000378:metabolism*; D004926:Escherichia coli / Q000737:chemistry; D006801:Humans; D019169:Jurkat Cells; D007700:Kinetics; D007711:Klebsiella pneumoniae / Q000737:chemistry; D008051:Lipid Bilayers / Q000737:chemistry / Q000378:metabolism; D008070:Lipopolysaccharides / Q000737:chemistry / Q000378:metabolism*; D008563:Membrane Lipids / Q000737:chemistry / Q000378:metabolism; D010713:Phosphatidylcholines / Q000737:chemistry / Q000378:metabolism; D010714:Phosphatidylethanolamines / Q000737:chemistry / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000378:metabolism; D019779:Salmonella enterica / Q000737:chemistry; D013050:Spectrometry, Fluorescence; D013109:Sphingomyelins / Q000737:chemistry / Q000378:metabolism; D013601:T-Lymphocytes / Q000378:metabolism*
D008051:Lipid Bilayers; D008070:Lipopolysaccharides; D008563:Membrane Lipids; D010713:Phosphatidylcholines; D010714:Phosphatidylethanolamines; D011956:Receptors, Cell Surface; D013109:Sphingomyelins; C483858:phosphatidylethanolamine; D002784:Cholesterol