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  • 1985-1989  (1)
  • 1980-1984  (2)
  • 1
    ISSN: 1432-072X
    Keywords: KDO mutant ; Lipid A intermediates ; Hexadecanoic acid substitution ; Biosynthesis of lipid A
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The extraction, purification and structural characterization of two lipid A precursors (Ia and Ib) differing only in one hexadecanoic acid are described. Both precursors were synthesized at elevated temperatures by a new mutant of Salmonella typhimurium (mutant Ts5) which is conditionally defective in synthesis of the 3-deoxy-d-manno-octulosonic acid region of lipopolysaccharides. Both precursors were purified by repeated phenol/chloroform/petroleum ether (PCP) extractions followed by thin layer chromatography. Teh precursor preparation was free of lipopolysaccharides and phospholipids and contained less than 0.1% protein. Structural analysis which included chemical degradation procedures as well as positive ion laser desorption (LDMS) mass spectroscopy of dephosphorylated lipid A precursors showed together that precursor Ia represents a diphosphorylated glucosamine disaccharide containing two ester, two amide-linked residues of 3-hydroxytetradecanoic acid and lacks the ester-linked dodecanoic, tetradecanoic and hexadecanoic acid as well as 3-deoxy-d-manno-octulosonic acid. Precursor Ib has the same basic structure as precursor Ia, but contains in addition one mol of hexadecanoic acid per mol disaccharide which is linked to the 3-hydroxy group of the amide-bound 3-hydroxy-tetradecanoic acid of the reducing, terminal glucosamine residue. The structure of precursor Ib supports the conclusion that hexadecanoic acid incorporation occurs at an early stage in lipid A biosynthesis prior to the attachment of 3-deoxy-d-manno-octulosonic acid and/or other polar substituents.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-1440
    Keywords: Lipid A ; Lipopolysaccharides ; Chemical structure ; Biological activity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Lipopolysaccharides are integral components of the outer membrane of Gram-negative bacteria and they participate in various membrane functions essential for bacterial growth and survival. Lipopolysaccharides also represent the endotoxins of Gram-negative bacteria and possibly play a role for the pathogenesis and manifestations of bacterial infections. These biological activities are mediated mainly by the lipid component of lipopolysaccharides, termed lipid A. Chemically, lipid A consists of a β1,6-linkedD-glucosamine disaccharide which carries substituted phosphoryl groups and a range ofD-3-hydroxy andD-3-acyloxyacyl residues, the latter being arranged in a hexagonal dense packing. A number of experimental data allow the conclusion that the highly ordered and compact lipid A structure confers stability to the outer membrane, renders it less permeable to lipophilic molecules and by providing a proper fluidity stabilizes the conformation of biologically active membrane proteins. For endotoxic activities of lipid A the polar substituents of phosphate residues are dispensable. The presence ofD-3-hydroxy (or acyloxy) acyl-groups, linked to the glucosamine disaccharide, however, seems to be of importance. Analyses of now available synthetic lipid A analogues are expected to allow a more precise characterization of substructures and conformations required for the expression of physiological functions and endotoxic activities of lipid A.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Chichester : Wiley-Blackwell
    Biological Mass Spectrometry 11 (1984), S. 132-141 
    ISSN: 0306-042X
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: The applicability and the present limitations of the laser microprobe mass analyser LAMMA®-500 as an instrument for the structural analysis of higher molecular weight, non-volatile, bio-organic compounds (≤2000 u) were investigated. For this purpose mass spectra of various synthetic and natural compounds representing cell wall components of Gram-negative bacteria, e.g. phospholipids and lipid A-like molecules were studied. In several cases these spectra exhibited relatively simple and interpretable patterns with a prominent quasi-molecular ion originating from alkali attachment. For one group of the compounds studied - synthetic lipid A-like molecules containing a phosphate moiety - the spectra were rather complicated and lacked pronounced quasi-molecular peaks. Possible reasons for this observation are discussed.
    Additional Material: 12 Ill.
    Type of Medium: Electronic Resource
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