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Häussler, S; Nimtz, M; Domke, T; Wray, V; Steinmetz, I.
Purification and characterization of a cytotoxic exolipid of Burkholderia pseudomallei.
Infect Immun. 1998; 66(4):1588-1593 Doi: 10.1128/IAI.66.4.1588-1593.1998 [OPEN ACCESS]
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Leading authors Med Uni Graz: Steinmetz Ivo
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Abstract:: Burkholderia pseudomallei is the causative agent of melioidosis, an infectious disease, which is increasingly recognized as an important public health problem in various tropical regions. This study describes the identification and characterization of a heat-stable extracellular toxin of B. pseudomallei. After cultivation of B. pseudomallei in liquid media, the heated cell-free supernatant was concentrated by ultrafiltration. The concentrate exhibited a cytotoxic and hemolytic activity which showed remarkable resistance against alkaline and acidic treatments. For further purification, reversed-phase chromatography using a fast-performance liquid chromatography system was performed. After elution with an acetonitrile gradient, a single cytotoxic and hemolytic peak was detected. Structural characterization of the toxin was performed by a combination of mass spectrometric and nuclear magnetic resonance spectroscopic techniques. A highly purified glycolipid, 2-O-alpha-L-rhamnopyranosyl-alpha-L-rhamnopyranosyl-beta-hydroxytetradec anoyl-beta-hydroxytetradecanoate (Rha-Rha-C14-C14), with a molecular mass of 762 Da was identified. The purified exolipid showed a time- and dose-dependent cytotoxic effect on phagocytic (HL60) and nonphagocytic (HeLa) cell lines. In addition, a time- and dose-dependent hemolysis of erythrocytes from various species was observed. The toxin structure makes a detergentlike action most probable. Interestingly, the cytotoxic and hemolytic activities of the glycolipid could be neutralized by albumin. Future studies will concentrate on the role of this exolipid as a virulence factor in the pathogenesis of melioidosis.
Find related publications in this database (using NLM MeSH Indexing): Animals -; Bacterial Toxins - chemistry; Bacterial Toxins - isolation & purification; Burkholderia pseudomallei - chemistry; Glycolipids - chemistry; Glycolipids - isolation & purification; HL-60 Cells -; HeLa Cells -; Hemolysis -; Humans -; Mice -; Sheep -