Structural analysis of the lipopolysaccharide from Chlamydophila psittaci strain 6BC.
The lipopolysaccaride of Chlamydophila psittaci 6BC was isolated from tissue culture-grown elementary bodies using a modified phenol/water procedure followed by extraction with phenol/chloroform/light petroleum. Compositional analyses indicated the presence of 3-deoxy-Dmanno-oct-2-ulosonic acid, GlcN, organic bound phosphate and fatty acids in a molar ratio of approximately 3. 3 : 2 : 1.8 : 4.6. Deacylated lipopolysaccharide was obtained after successive microscale treatment with hydrazine and potassium hydroxide, and was then separated by high performance anion-exchange chromatography into two major fractions, the structures of which were determined by 600 MHz NMR spectroscopy as alpha-Kdo-(2-->8)-alpha-Kdo-(2-->4)-alpha-Kdo-(2-->6)-beta-D-GlcpN -(1 -->6)-alpha-D-GlcpN 1,4'-bisphosphate and alpha-Kdo-(2-->4)-[alpha-Kdo-(2-->8)]-alpha-Kdo-(2-->4)-alpha-Kdo-(2- ->6)-beta-D-GlcpN-(1-->6)-alpha-D-GlcpN 1,4'-bisphosphate. The distribution of fatty acids in lipid A was determined by compositional analyses and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry experiments on lipid A and de-O-acylated lipid A. It was shown that the carbohydrate backbone of lipid A is replaced by a complex mixture of fatty acids, including long-chain and branched (R)-configured 3-hydroxy fatty acids, the latter being exclusively present in an amide linkage.
Rund
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Divisions of Medical and Biochemical Microbiology and Biophysics, Research Center Borstel, Center for Medicine and Biosciences, Germany.
Lindner
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B
Brade
H
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Holst
O
O
D000818:Animals; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002460:Cell Line; D002691:Chlamydophila psittaci / Q000737:chemistry*; D002725:Chloroform / Q000737:chemistry; D002852:Chromatography, Ion Exchange; D002855:Chromatography, Thin Layer; D006834:Hydrazines / Q000737:chemistry; D006878:Hydroxides / Q000494:pharmacology; D008050:Lipid A / Q000737:chemistry / Q000302:isolation & purification; D008070:Lipopolysaccharides / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy; D051379:Mice; D008969:Molecular Sequence Data; D019800:Phenol / Q000737:chemistry; D017680:Potassium Compounds / Q000494:pharmacology; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013400:Sugar Acids / Q000737:chemistry; D013997:Time Factors; D014867:Water / Q000737:chemistry
D006834:Hydrazines; D006878:Hydroxides; D008050:Lipid A; D008070:Lipopolysaccharides; D017680:Potassium Compounds; D013400:Sugar Acids; D014867:Water; C002532:2-keto-3-deoxyoctonate; C029424:hydrazine; D019800:Phenol; D002725:Chloroform; C029943:potassium hydroxide