A special acyl carrier protein for transferring long hydroxylated fatty acids to lipid A in Rhizobium.

The Journal of Biological Chemistry
K A BrozekChristian R H Raetz

Abstract

Lipid A, the hydrophobic anchor of lipopolysaccharides in the outer membranes of Gram-negative bacteria, varies in structure among different Rhizobiaceae. The Rhizobium meliloti lipid A backbone, like that of Escherichia coli, is a beta1'-6-linked glucosamine disaccharide that is phosphorylated at positions 1 and 4'. Rhizobium leguminosarum lipid A lacks both phosphates, but contains aminogluconate in place of the proximal glucosamine 1-phosphate, and galacturonic acid instead of the 4'-phosphate. A peculiar feature of the lipid As of all Rhizobiaceae is acylation with 27-hydroxyoctacosanoic acid, a long hydroxylated fatty acid not found in E. coli. We now describe an in vitro system, consisting of a membrane enzyme and a cytosolic acyl donor from R. leguminosarum, that transfers 27-hydroxyoctacosanoic acid to (Kdo)2-lipid IVA, a key lipid A precursor common to both E. coli and R. leguminosarum. The 27-hydroxyoctacosanoic acid moiety was detected in the lipid product by mass spectrometry. The membrane enzyme required the presence of Kdo residues in the acceptor substrate for activity. The cytosolic acyl donor was purified from wild-type R. leguminosarum using the acylation of (Kdo)2-[4'-32P]-lipid IVA as the assay. Amino-termin...Continue Reading

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Citations

Dec 13, 2003·Carbohydrate Research·Martine Caroff, Doris Karibian
May 23, 2002·The Journal of Biological Chemistry·Shib Sankar BasuChristian R H Raetz
Jan 18, 2003·The Journal of Biological Chemistry·Nanette L S Que-GewirthChristian R H Raetz
Jan 18, 2003·The Journal of Biological Chemistry·Nanette L S Que-GewirthChristian R H Raetz
Jul 19, 2003·The Journal of Biological Chemistry·Mark J KarbarzChristian R H Raetz
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