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Lipopolysaccharide ofRhodospirillum salinarum 40: structural studies on the core and lipid A region

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Abstract

The structural elucidation of lipid A of the cell wall lipopolysaccharide (LPS) ofRhodospirillum salinarum 40 by chemical methods and laser desorption mass spectrometry revealed the presence of a mixed lipid A composed of three different 1,4 bisphosphorylated β(1→6)-linked backbone hexosaminyl-hexosamine disaccharides, i.e. those composed of GlCN→GlcN, 2,3-diamino-2,3-dideoxy-d-Glc-(DAG)→DAG, and DAG→GlcN. Lipid A ofR. salinarum contained preferentially 3-OH-18:0 and 3-OH-14:0 as amide-linked andcisΔ11-18:1 and c19:0 as ester-linked fatty acids. The mass spectra of the liberated acyl-oxyacyl residues proved the concomitant presence of 3-O-(cisΔ11-18:1)-18:0 and 3-O-(c19:0)-14:0 as the predominating diesters in this mixed lipid A. The glycosidically linked and the ester-linked phosphate groups of the backbone disaccharide were neither substituted by ethanolamine phosphorylethanolamine, nor by 4-amino-4-deoxy-l-arabinose, in contrast to most of the enterobacterial lipid As. In the core oligosaccharide fraction, a HexA (1→4)HexA(1→5)Kdo-trisaccharide was identified by methylation analysis. The terminal HexA (hexuronic acid) is possibly 4-OMe-GalA, a component described here as an LPS constituent for the first time. LPS ofR. salinarum showed a lethality in C57BL/10 ScSN (LPS-responder)-mice) of an order of 10−1–10−2 of that reported forSalmonella abortus equi LPS, and it was also capable of inducing TNFα and IL6 in macrophages of C57BL/10ScSN mice.

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Abbreviations

AAS :

Atomic absorption spectroscopy

c19:0 cis-11, 12:

Methylene-octadecanoic acid

DAG :

2,3-Diamino-2,3-dideoxy-d-glucose

DMDS :

Dimethyldisulfide

DOC-PAGE :

Deoxycholate-polyacrylamide gel electrophoresis

EI-MS :

Electron impact mass spectrometry

GalA :

Galacturonic acid

GlcA :

Glucuronic acid

GC-MS :

Combined gas liquid chromatography-mass spectrometry

GlcN :

d-Glucosamine

HexA :

Hexuronic acid

IL1 :

Interleukin 1

IL6 :

Interleukin 6

Kdo :

3-Deoxy-d-manno-octulosonate

LD-MS :

Laser desorption mass spectrometry

LPS :

Lipopolysaccharide

MTT :

3-(4,5-Dimethlythiazol-2-yl)-2,5-diphenyltetrazoliumbromide

4-OMe :

GalA 4-O-methylgalacturonic acid

PITC :

Phenyl isothiocyanate

TNFα :

Tumor necrosis factor α

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Authors and Affiliations

  1. Max-Planck-Institut für Immunbiologie, Stübeweg 51, D-79108, Freiburg i. Br., Germany

    Heike Rau, Marina Freudenberg & Hubert Mayer

  2. Forschungsinstitut Borstel, Parkallee 22, D-23845, Borstel, Germany

    Ulrich Seydel

  3. Institut für Biologie II, Mikrobiologie, Schänzlestrasse 1, D-79104, Freiburg i. Br., Germany

    Jürgen Weckesser

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  1. Heike Rau
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  2. Ulrich Seydel
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  4. Jürgen Weckesser
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Rau, H., Seydel, U., Freudenberg, M. et al. Lipopolysaccharide ofRhodospirillum salinarum 40: structural studies on the core and lipid A region. Arch. Microbiol. 164, 280–289 (1995). https://doi.org/10.1007/BF02529962

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  • DOI: https://doi.org/10.1007/BF02529962

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