title: Biophysical characterization of triacyl monosaccharide lipid a partial structures in relation to bioactivity.
abstract: Synthetic triacyl glucosamine monosaccharide lipid A part structures corresponding to the non-reducing moiety of enterobacterial lipid A with an acyloxyacyl chain linked to position 3 of the glucosamine and an unbranched chain linked to position 2 (group 1) and vice versa (group 2) were analyzed biophysically: Fourier-transform infrared spectroscopy was performed to characterize the gel-to-liquid crystalline phase transition, the phosphate band contour, and the orientation of the glucosamine with respect to the membrane surface. Small-angle x-ray diffraction was applied for the elucidation of the supramolecular aggregate structure and, with that, of the molecular shape. With fluorescence resonance energy transfer the lipopolysaccharide-binding protein (LBP)-mediated intercalation of the lipid A partial structures into phospholipid liposomes was monitored. The physical data clearly exhibit a classification of the synthetic compounds into two groups: group 1 compounds have sharp phase transitions, indicating dense acyl chain packing and an inclination of the glucosamine backbone with respect to the membrane surface of 30 degrees with the phosphate buried in the membrane. Group 2 compounds have a very broad phase transition, indicating poorly packed acyl chains, and an inclination of -30 degrees with the phosphate group sticking outward. For the first group unilamellar phases are observed superimposed by a non-lamellar structure, and for the second one only multilamellar aggregate structures. The cytokine-inducing capacity in human mononuclear cells is relatively high for the first group and low or absent for the second group. Based on these data a model of the intra and intermolecular conformations is proposed which also extends the concept of "endotoxic conformation."
authors.affiliation: Forschungszentrum Borstel, Division of Biophysics, D-23845 Borstel, Germany. kbranden@fz-borstel.de
authors.forename: Klaus
authors.initials: K
authors.lastname: Brandenburg
authors.forename: Motohiro
authors.initials: M
authors.lastname: Matsuura
authors.forename: Holger
authors.initials: H
authors.lastname: Heine
authors.forename: Mareike
authors.initials: M
authors.lastname: Müller
authors.forename: Makato
authors.initials: M
authors.lastname: Kiso
authors.forename: Hideharu
authors.initials: H
authors.lastname: Ishida
authors.forename: Michel H J
authors.initials: MH
authors.lastname: Koch
authors.forename: Ulrich
authors.initials: U
authors.lastname: Seydel
mesh_terms: D000818:Animals; D055592:Biophysical Phenomena; D001703:Biophysics; D016466:CHO Cells; D006224:Cricetinae; D016207:Cytokines / Q000096:biosynthesis; D004305:Dose-Response Relationship, Drug; D029721:Drosophila Proteins; D005944:Glucosamine / Q000737:chemistry; D006017:Glycolipids / Q000378:metabolism; D006737:Horseshoe Crabs / Q000378:metabolism; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000378:metabolism; D008027:Light; D008050:Lipid A / Q000737:chemistry*; D018950:Lipopolysaccharide Receptors / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000378:metabolism; D008954:Models, Biological; D008956:Models, Chemical; D009005:Monosaccharides / Q000737:chemistry*; D011487:Protein Conformation; D011956:Receptors, Cell Surface / Q000378:metabolism; D012542:Scattering, Radiation; D017550:Spectroscopy, Fourier Transform Infrared; D013696:Temperature; D051193:Toll-Like Receptors; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism; D014961:X-Ray Diffraction
chemical_list: D016207:Cytokines; D029721:Drosophila Proteins; D006017:Glycolipids; D008050:Lipid A; D018950:Lipopolysaccharide Receptors; D008562:Membrane Glycoproteins; D009005:Monosaccharides; D011956:Receptors, Cell Surface; D051193:Toll-Like Receptors; D014409:Tumor Necrosis Factor-alpha; D005944:Glucosamine