PMID:11717579

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Citation

Gronow, S, Oertelt, C, Ervelä, E, Zamyatina, A, Kosma, P, Skurnik, M and Holst, O (2001) Characterization of the physiological substrate for lipopolysaccharide heptosyltransferases I and II. J. Endotoxin Res. 7:263-70

Abstract

L-Glycero-D-manno-heptopyranose is a characteristic compound of many lipopolysaccharide (LPS) core structures of Gram-negative bacteria. In Escherichia coli two heptosyltransferases, namely WaaC and WaaF, are known to transfer L-glycero-D-manno-heptopyranose to Re-LPS and Rd(2)-LPS, respectively. It had been proposed that both reactions involve ADPL-glycero-D-manno-heptose as a sugar donor; however, the structure of this nucleotide sugar had never been completely elucidated. In the present study, ADPL-glycero-D-manno-heptose was isolated from a heptosyltransferase-deficient E. coli mutant, and its structure was determined by nuclear magnetic resonance spectroscopy and matrix-assisted laser-desorption/ionization time-of-flight mass spectrometry as ADPL-glycero-beta-D-manno-heptopyranose. This compound represented the sole constituent of the bacterial extract that was accepted as a sugar donor by heptosyltransferases I and II in vitro.

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Keywords

Adenosine Diphosphate Sugars/metabolism; Escherichia coli/enzymology; Glycosyltransferases/metabolism; Heptoses/chemistry; Heptoses/metabolism; Nuclear Magnetic Resonance, Biomolecular; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Substrate Specificity/physiology

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