Stable isotopic labelling-assisted untargeted metabolic profiling reveals novel conjugates of the mycotoxin deoxynivalenol in wheat

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• 作者：Bernhard Kluger (1)
  Christoph Bueschl (1)
  Marc Lemmens (2)
  Franz Berthiller (1)
  Georg H?ubl (3)
  Günther Jaunecker (3)
  Gerhard Adam (4)
  Rudolf Krska (1)
  Rainer Schuhmacher (1)
  
• 关键词：Metabolisation ; Xenobiotics ; Stable isotopic labelling ; Deoxynivalenol ; Liquid chromatography–high resolution mass spectrometry ; Mycotoxin conjugate
• 刊名：Analytical and Bioanalytical Chemistry
• 出版年：2013
• 出版时间：June 2013
• 年：2013
• 卷：405
• 期：15
• 页码：5031-5036
• 全文大小：248KB
• 参考文献：1. Goodacre R, Vaidyanathan S, Dunn WB, Harrigan GG, Kell DB (2004) Metabolomics by numbers: acquiring and understanding global metabolite data. Trends Biotechnol 22(5):245-52 CrossRef
  2. Patterson AD, Gonzalez FJ, Idle JR (2010) Xenobiotic metabolism: a view through the metabolometer. Chem Res Toxicol 23(5):851-60. doi:10.1021/tx100020p CrossRef
  3. Chen C, Gonzalez FJ, Idle JR (2007) LC-MS-based metabolomics in drug metabolism. Drug Metab Rev 39(2-):581-97. doi:10.1080/03602530701497804 CrossRef
  4. Patti GJ, Yanes O, Siuzdak G (2012) Innovation: metabolomics: the apogee of the omics trilogy. Nat Rev Mol Cell Biol 13(4):263-69 CrossRef
  5. Birkemeyer C, Luedemann A, Wagner C, Erban A, Kopka J (2005) Metabolome analysis: the potential of in vivo labeling with stable isotopes for metabolite profiling. Trends Biotechnol 23(1):28-3 CrossRef
  6. Hegeman AD, Schulte CF, Cui Q, Lewis IA, Huttlin EL, Eghbalnia H, Harms AC, Ulrich EL, Markley JL, Sussman MR (2007) Stable isotope assisted assignment of elemental compositions for metabolomics. Anal Chem 79(18):6912-921. doi:10.1021/ac070346t CrossRef
  7. Giavalisco P, Hummel J, Lisec J, Inostroza AC, Catchpole G, Willmitzer L (2008) High-resolution direct infusion-based mass spectrometry in combination with whole 13c metabolome isotope labeling allows unambiguous assignment of chemical sum formulas. Anal Chem 80(24):9417-425. doi:10.1021/ac8014627 CrossRef
  8. Giavalisco P, K?hl K, Hummel J, Seiwert B, Willmitzer L (2009) 13C isotope-labeled metabolomes allowing for improved compound annotation and relative quantification in liquid chromatography–mass spectrometry-based metabolomic research. Anal Chem 81(15):6546-551. doi:10.1021/ac900979e CrossRef
  9. Giavalisco P, Li Y, Matthes A, Eckhardt A, Hubberten H-M, Hesse H, Segu S, Hummel J, K?hl K, Willmitzer L (2011) Elemental formula annotation of polar and lipophilic metabolites using 13C, 15 N and 34S isotope labelling, in combination with high-resolution mass spectrometry. Plant J 68(2):364-76. doi:10.1111/j.1365-313X.2011.04682.x CrossRef
  10. Hiller K, Metallo C, Stephanopoulos G (2011) Elucidation of cellular metabolism via metabolomics and stable-isotope assisted metabolomics. Curr Pharm Biotechnol 12(7):1075-086 CrossRef
  11. Athersuch TJ, Nicholson JK, Wilson ID (2007) Isotopic enrichment enhancement in metabonomic analysis of UPLC-MS data sets. J Label Compd Radiopharm 50(5-):303-07. doi:10.1002/jlcr.1217 CrossRef
  12. Bueschl C, Kluger B, Berthiller F, Lirk G, Winkler S, Krska R, Schuhmacher R (2012) MetExtract: a new software tool for the automated comprehensive extraction of metabolite-derived LC/MS signals in metabolomics research. Bioinformatics. doi:10.1093/bioinformatics/bts012
  13. Pestka J (2010) Deoxynivalenol: mechanisms of action, human exposure, and toxicological relevance. Arch Toxicol 84(9):663-79. doi:10.1007/s00204-010-0579-8 CrossRef
  14. Walter S, Nicholson P, Doohan FM (2010) Action and reaction of host and pathogen during / Fusarium head blight disease. New Phytol 185(1):54-6. doi:10.1111/j.1469-8137.2009.03041.x CrossRef
  15. Lemmens M, Scholz U, Berthiller F, Dall'Asta C, Koutnik A, Schuhmacher R, Adam G, Buerstmayr H, Mesterházy á, Krska R, Ruckenbauer P (2005) The ability to detoxify the mycotoxin deoxynivalenol colocalizes with a major quantitative trait locus for fusarium head blight resistance in wheat. Mol Plant-Microbe Interact 18(12):1318-324. doi:10.1094/mpmi-18-1318 CrossRef
  16. Proctor RH, Hohn TM, McCormick SP (1995) Reduced virulence of / Gibberella zeae caused by disruption of a trichothecene toxin biosynthetic gene. Mol Plant-Microbe Interact 8(4):593-01 CrossRef
  17. Poppenberger B, Berthiller F, Lucyshyn D, Sieberer T, Schuhmacher R, Krska R, Kuchler K, Gl?ssl J, Luschnig C, Adam G (2003) Detoxification of the fusarium mycotoxin deoxynivalenol by a UDP-glucosyltransferase from / Arabidopsis thaliana. J Biol Chem 278(48):47905-7914. doi:10.1074/jbc.M307552200 CrossRef
  18. Berthiller F, Dall'Asta C, Schuhmacher R, Lemmens M, Adam G, Krska R (2005) Masked mycotoxins: determination of a deoxynivalenol glucoside in artificially and naturally contaminated wheat by liquid chromatography–tandem mass spectrometry. J Agric Food Chem 53(9):3421-425. doi:10.1021/jf047798g CrossRef
  19. De Vos RCH, Moco S, Lommen A, Keurentjes JJB, Bino RJ, Hall RD (2007) Untargeted large-scale plant metabolomics using liquid chromatography coupled to mass spectrometry. Nat Protocols 2(4):778-91 CrossRef
  20. Levsen K, Schiebel H-M, Behnke B, D?tzer R, Dreher W, Elend M, Thiele H (2005) Structure elucidation of phase II metabolites by tandem mass spectrometry: an overview. J Chromatogr A 1067(1-):55-2
  21. Berthiller F, Schuhmacher R, Adam G, Krska R (2009) Formation, determination and significance of masked and other conjugated mycotoxins. Anal Bioanal Chem 395(5):1243-252. doi:10.1007/s00216-009-2874-x CrossRef
  22. Gardiner SA, Boddu J, Berthiller F, Hametner C, Stupar RM, Adam G, Muehlbauer GJ (2010) Transcriptome analysis of the barley–deoxynivalenol interaction: evidence for a role of glutathione in deoxynivalenol detoxification. Mol Plant-Microbe Interact 23(7):962-76. doi:10.1094/mpmi-23-7-0962 CrossRef
  23. Schw?bel JAH, Madden JC, Cronin MTD (2010) Examination of Michael addition reactivity towards glutathione by transition-state calculations. SAR QSAR Environ Res 21(7-):693-10. doi:10.1080/1062936x.2010.528943 CrossRef
  24. Marrs KA (1996) The functions and regulation of glutathione s-transferases in plants. Annu Rev Plant Physiol Plant Mol Biol 47(1):127-58. doi:10.1146/annurev.arplant.47.1.127 CrossRef
  
• 作者单位：Bernhard Kluger (1)
  Christoph Bueschl (1)
  Marc Lemmens (2)
  Franz Berthiller (1)
  Georg H?ubl (3)
  Günther Jaunecker (3)
  Gerhard Adam (4)
  Rudolf Krska (1)
  Rainer Schuhmacher (1)
  
  1. Center for Analytical Chemistry, Department for Agrobiotechnology (IFA-Tulln), University of Natural Resources and Life Sciences (BOKU), Vienna, Konrad-Lorenz-Str. 20, 3430, Tulln, Austria
  2. Institute for Biotechnology in Plant Production, Department for Agrobiotechnology (IFA-Tulln), University of Natural Resources and Life Sciences (BOKU), Vienna, Konrad-Lorenz-Str. 20, 3430, Tulln, Austria
  3. Romer Labs Diagnostic GmbH, Technopark 1, 3430, Tulln, Austria
  4. Department of Applied Genetics and Cell Biology, University of Natural Resources and Life Sciences (BOKU), Vienna, Konrad-Lorenz-Str. 24, 3430, Tulln, Austria
  
• ISSN：1618-2650

文摘

An untargeted screening strategy for the detection of biotransformation products of xenobiotics using stable isotopic labelling (SIL) and liquid chromatography–high resolution mass spectrometry (LC-HRMS) is reported. The organism of interest is treated with a mixture of labelled and non-labelled precursor and samples are analysed by LC-HRMS. Raw data are processed with the recently developed MetExtract software for the automated extraction of corresponding peak pairs. The SIL-assisted approach is exemplified by the metabolisation of the Fusarium mycotoxin deoxynivalenol (DON) in planta. Flowering ears were inoculated with 100?μg of a 1-- (v/v) mixture of non-labelled and fully labelled DON. Subsequent sample preparation, LC-HRMS measurements and data processing revealed a total of 57 corresponding peak pairs, which originated from ten metabolites. Besides the known DON and DON-3-glucoside, which were confirmed by measurement of authentic standards, eight further DON-biotransformation products were found by the untargeted screening approach. Based on a mass deviation of less than ±5?ppm and MS/MS measurements, one of these products was annotated as DON-glutathione (GSH) conjugate, which is described here for the first time for wheat. Our data further suggest that two DON-GSH-related metabolites, the processing products DON-S-cysteine and DON-S-cysteinyl-glycine and five unknown DON conjugates were formed in planta. Future MS/MS measurements shall reveal the molecular structures of the detected conjugates in more detail.