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Journal of Endotoxin Research
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Characterization of the physiological substrate for lipopolysaccharide heptosyltransferases I and II

Sabine Gronow

Medical and Biochemical Microbiology, Research Center Borstel, Germany

Clemens Oertelt

Analytical Biochemistry, Research Center Borstel, Germany

Elise Ervelä

Department of Medical Biochemistry, University of Turku, Finland

Alla Zamyatina

Department of Chemistry, University of Agricultural Sciences, Vienna, Austria

Paul Kosma

Department of Chemistry, University of Agricultural Sciences, Vienna, Austria

Mikael Skurnik

Department of Medical Biochemistry, University of Turku, Finland

Otto Holst

Analytical Biochemistry, Research Center Borstel, Germany, oholst{at}fz-borstel.de

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-ß-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 .

Journal of Endotoxin Research, Vol. 7, No. 4, 263-270 (2001)
DOI: 10.1177/09680519010070040701
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