Dynamic tracing of sugar metabolism reveals the mechanisms of action of synthetic sugar analogs

Monique Scherpenzeel; Federica Conte; Christian Büll; Angel Ashikov; Esther Hermans; Anke Willems; Walinka 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

doi:10.1093/glycob/cwab106

Dynamic tracing of sugar metabolism reveals the mechanisms of action of synthetic sugar analogs

Monique Scherpenzeel, Federica Conte, Christian Büll, Angel Ashikov, Esther Hermans, Anke Willems, Walinka 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^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › Academic › peer-review

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 ultrahigh performance liquid chromatography-triple quadrupole mass spectrometry 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 3F(ax)-NeuNAc by incubation of B16-F10 melanoma cells with N-acetyl-D-[UL-(13)C6]glucosamine revealed full depletion of endogenous ManNAc 6-phosphate and CMP-NeuNAc within 24 h. 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.

Original language	English
Pages (from-to)	239-250
Number of pages	12
Journal	Glycobiology
Volume	32
Issue number	3
Early online date	25 Oct 2021
DOIs	https://doi.org/10.1093/glycob/cwab106
Publication status	Published - 30 Mar 2022

Keywords

BIOSYNTHESIS
GLYCOSYLATION
IDENTIFICATION
INHIBITION
MASS-SPECTROMETRY
MUTATIONS
NUCLEOTIDE SUGARS
SIALIC-ACID
SIALYLATION
TARGET
fluoro sialic acid
glycosylation
metabolic oligosaccharide engineering
sugar metabolism
synthetic sugar analog

Access to Document

10.1093/glycob/cwab106Licence: CC BY

Cite this

Scherpenzeel, M., Conte, F., Büll, C., Ashikov, A., Hermans, E., Willems, A., Tol, W., Kragt, E., Noga, M., Moret, E. E., Heise, T., Langereis, J. D., Rossing, E., Zimmermann, M., Rubio-Gozalbo, M. E., de Jonge, M. I., Adema, G. J., Zamboni, N., Boltje, T., & Lefeber, D. J. (2022). Dynamic tracing of sugar metabolism reveals the mechanisms of action of synthetic sugar analogs. Glycobiology, 32(3), 239-250. https://doi.org/10.1093/glycob/cwab106

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title = "Dynamic tracing of sugar metabolism reveals the mechanisms of action of synthetic sugar analogs",

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 ultrahigh performance liquid chromatography-triple quadrupole mass spectrometry 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 3F(ax)-NeuNAc by incubation of B16-F10 melanoma cells with N-acetyl-D-[UL-(13)C6]glucosamine revealed full depletion of endogenous ManNAc 6-phosphate and CMP-NeuNAc within 24 h. 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.",

keywords = "BIOSYNTHESIS, GLYCOSYLATION, IDENTIFICATION, INHIBITION, MASS-SPECTROMETRY, MUTATIONS, NUCLEOTIDE SUGARS, SIALIC-ACID, SIALYLATION, TARGET, fluoro sialic acid, glycosylation, metabolic oligosaccharide engineering, sugar metabolism, synthetic sugar analog",

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note = "{\textcopyright} The Author(s) 2021. Published by Oxford University Press.",

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doi = "10.1093/glycob/cwab106",

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Scherpenzeel, M, Conte, F, Büll, C, Ashikov, A, Hermans, E, Willems, A, Tol, W, Kragt, E, Noga, M, Moret, EE, Heise, T, Langereis, JD, Rossing, E, Zimmermann, M, Rubio-Gozalbo, ME, de Jonge, MI, Adema, GJ, Zamboni, N, Boltje, T & Lefeber, DJ 2022, 'Dynamic tracing of sugar metabolism reveals the mechanisms of action of synthetic sugar analogs', Glycobiology, vol. 32, no. 3, pp. 239-250. https://doi.org/10.1093/glycob/cwab106

TY - JOUR

T1 - Dynamic tracing of sugar metabolism reveals the mechanisms of action of synthetic sugar analogs

AU - Scherpenzeel, Monique

AU - Conte, Federica

AU - Büll, Christian

AU - Ashikov, Angel

AU - Hermans, Esther

AU - Willems, Anke

AU - Tol, Walinka

AU - Kragt, Else

AU - Noga, Marek

AU - Moret, Ed E

AU - Heise, Torben

AU - Langereis, Jeroen D

AU - Rossing, Emiel

AU - Zimmermann, Michael

AU - Rubio-Gozalbo, M Estela

AU - de Jonge, Marien I

AU - Adema, Gosse J

AU - Zamboni, Nicola

AU - Boltje, Thomas

AU - Lefeber, Dirk J

N1 - © The Author(s) 2021. Published by Oxford University Press.

PY - 2022/3/30

Y1 - 2022/3/30

N2 - 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 ultrahigh performance liquid chromatography-triple quadrupole mass spectrometry 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 3F(ax)-NeuNAc by incubation of B16-F10 melanoma cells with N-acetyl-D-[UL-(13)C6]glucosamine revealed full depletion of endogenous ManNAc 6-phosphate and CMP-NeuNAc within 24 h. 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.

AB - 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 ultrahigh performance liquid chromatography-triple quadrupole mass spectrometry 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 3F(ax)-NeuNAc by incubation of B16-F10 melanoma cells with N-acetyl-D-[UL-(13)C6]glucosamine revealed full depletion of endogenous ManNAc 6-phosphate and CMP-NeuNAc within 24 h. 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.

KW - BIOSYNTHESIS

KW - GLYCOSYLATION

KW - IDENTIFICATION

KW - INHIBITION

KW - MASS-SPECTROMETRY

KW - MUTATIONS

KW - NUCLEOTIDE SUGARS

KW - SIALIC-ACID

KW - SIALYLATION

KW - TARGET

KW - fluoro sialic acid

KW - glycosylation

KW - metabolic oligosaccharide engineering

KW - sugar metabolism

KW - synthetic sugar analog

U2 - 10.1093/glycob/cwab106

DO - 10.1093/glycob/cwab106

M3 - Article

C2 - 34939087

SN - 0959-6658

VL - 32

SP - 239

EP - 250

JO - Glycobiology

JF - Glycobiology

IS - 3

ER -