1. Dynamic tracing of sugar metabolism reveals the mechanisms of action of synthetic sugar analogs
- Author
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Monique van Scherpenzeel, Federica Conte, Christian Büll, Angel Ashikov, Esther Hermans, Anke Willems, Walinka van Tol, Else Kragt, Marek Noga, Ed E Moret, Torben Heise, Jeroen D Langereis, Emiel Rossing, Michael Zimmermann, M Estela Rubio-Gozalbo, Marien I de Jonge, Gosse J Adema, Nicola Zamboni, Thomas Boltje, Dirk J Lefeber, Kindergeneeskunde, MUMC+: MA Medische Staf Kindergeneeskunde (9), and RS: GROW - R4 - Reproductive and Perinatal Medicine
- Subjects
glycosylation ,Cancer development and immune defence Radboud Institute for Molecular Life Sciences [Radboudumc 2] ,INHIBITION ,Stress-related disorders Donders Center for Medical Neuroscience [Radboudumc 13] ,lnfectious Diseases and Global Health Radboud Institute for Molecular Life Sciences [Radboudumc 4] ,Synthetic Organic Chemistry ,synthetic sugar analog ,Biochemistry ,130 000 Cognitive Neurology & Memory ,sugar metabolism ,BIOSYNTHESIS ,fluoro sialic acid ,metabolic oligosaccharide engineering ,SIALIC-ACID ,Glucosamine ,IDENTIFICATION ,MUTATIONS ,SIALYLATION ,Fluoro sialic acid ,Metabolic Disorders Radboud Institute for Molecular Life Sciences [Radboudumc 6] ,MASS-SPECTROMETRY ,Disorders of movement Donders Center for Medical Neuroscience [Radboudumc 3] ,carbohydrates (lipids) ,TARGET ,Cytidine Monophosphate N-Acetylneuraminic Acid ,Carbohydrate Metabolism ,Sugars ,NUCLEOTIDE SUGARS ,Chromatography, Liquid - Abstract
Synthetic sugar analogs are widely applied in metabolic oligosaccharide engineering (MOE) and as novel drugs to interfere with glycoconjugate biosynthesis. However, mechanistic insights on their exact cellular metabolism over time are mostly lacking. We combined ion-pair UHPLC-QqQ mass spectrometry using tributyl- and triethylamine buffers for sensitive analysis of sugar metabolites in cells and organisms and identified low abundant nucleotide sugars, such as UDP-arabinose in human cell lines and CMP-sialic acid (CMP-NeuNAc) in Drosophila. Furthermore, MOE revealed that propargyloxycarbonyl (Poc) labeled ManNPoc was metabolized to both CMP-NeuNPoc and UDP-GlcNPoc. Finally, time-course analysis of the effect of antitumor compound 3Fax-NeuNAc by incubation of B16-F10 melanoma cells with N-acetyl-D-[UL-13C6]glucosamine revealed full depletion of endogenous ManNAc 6-phosphate and CMP-NeuNAc within 24 hour. Thus, dynamic tracing of sugar metabolic pathways provides a general approach to reveal time-dependent insights into the metabolism of synthetic sugars, which is important for the rational design of analogs with optimized effects., Glycobiology, 32 (3), ISSN:0959-6658
- Published
- 2022