Detergent-accelerated hydrolysis of bacterial endotoxins and determination of the anomeric configuration of the glycosyl phosphate present in the “Isolated lipid A” fragment of the Bordetella pertussis endotoxin
✍ Scribed by Martine Caroff; Angelika Tacken; Ladislas Szabó
- Publisher
- Elsevier Science
- Year
- 1988
- Tongue
- English
- Weight
- 707 KB
- Volume
- 175
- Category
- Article
- ISSN
- 0008-6215
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✦ Synopsis
Due to the formation of micelles, severance of the hydrophilic (poly-or oligosaccharide) and hydrophobic ("Lipid A") domains of bacterial lipopolysaccharides at pH 3.4 or 4.5 and 100" is slow and sometimes does not proceed at all; partially degraded fragments are usually formed. At pH 3.4 (100') in aqueous 1% sodium dodecylsulphate (SDS), both lipopolysaccharides of the Bordetellu pertussis endotoxin are cleaved within 20-30 min, but 80% of the glycosidically bound phosphate present in the hydrophobic domain is lost. Other endotoxins behave similarly. At pH 4.5 (100") and in the absence of detergent, hydrolysis of the glycosidic bonds of 3-deoxy-D-manno-2-octulosonic acid residues of the B. pertussis endotoxin is negligible but, in aqueous 1% SDS, severance of the two regions of LPS-1 is complete within 1 h (that of LPS-2 requires 3-4 h), and the glycosidically bound phosphate of the isolated hydrophobic region is preserved. Comparison of the rate of acid-catalysed hydrolysis of the glycosidically bound phosphate present in this "isolated Lipid A" preparation with that of 2-deoxy-2-[(3R)-3-hydroxytetradecanamidol-a-and -/3-D-glucopyranose l-phosphates established that the former l-phosphate was the (Y anomer. Salmonella minnesota 9595 (made up of two1 units of 3-deoxy-D-manno-2-*Dedicated to Professor Rezsd Bognk in the year of his 75th birthday.