Rapid analysis of nucleotide-activated sugars by high-performance liquid chromatography coupled with diode-array detection, electrospray ionization mass spectrometry and nuclear magnetic resonance
| Autor: | Emerson Ferreira Queiroz, Kurt Hostettmann, Michael Ramm, Jean-Luc Wolfender, Matthias Hamburger |
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| Rok vydání: | 2004 |
| Předmět: |
Spectrometry
Mass Electrospray Ionization Electrospray Magnetic Resonance Spectroscopy Electrospray ionization ANTIGEN Carbohydrates Analytical chemistry Mannose LIPOPOLYSACCHARIDE Mass spectrometry Nucleotide sugar Biochemistry High-performance liquid chromatography Analytical Chemistry chemistry.chemical_compound GDP-MANNOSE SOLVENT SUPPRESSION BIOSYNTHESIS Nucleotide Chromatography High Pressure Liquid chemistry.chemical_classification SPECTROSCOPY Chromatography Nucleotides Organic Chemistry PSEUDOMONAS-AERUGINOSA General Medicine GENE chemistry ESCHERICHIA-COLI ANIGOZANTHOS-PREISSII Guanosine diphosphate mannose |
| Zdroj: | Journal of Chromatography. A, Vol. 1034, No 1-2 (2004) pp. 139-148 |
| ISSN: | 0021-9673 |
| DOI: | 10.1016/j.chroma.2004.02.023 |
| Popis: | A generally applicable method for HPLC analysis of sugar nucleotides was established. Separation was achieved using ion-pair chromatography on a reversed-phase column. Ion-pair reagents were selected and various parameters optimized with respect to separation of I I of the most important sugar nucleotides and compatibility with on-line detection by electrospray ionization MS and NMR. The method was applied to the on-line analysis of the GDP-D-mannose-4,6-dehydratase (Gmd) and GDP-4-keto-6-deoxy-D-mannose reductase (Rmd) catalyzed conversion of GDP-D-mannose to GDP-D-rhamnose. By LC-NMR, the intermediate product of the reaction was shown to be a mixture of GDP-4-keto-6-deoxy-D-mannose and GDP-3-keto-6-deoxy-D-mannose. Nucleotide co-factors of enzymatic reactions such as ATP and NADH did not interfere with the analysis of nucleotide-activated sugars. (C) 2004 Elsevier B.V. All rights reserved. |
| Databáze: | OpenAIRE |
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