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Sulphur tracer experiments in laboratory animals using 34S-labelled yeast

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We have evaluated the use of 34S-labelled yeast to perform sulphur metabolic tracer experiments in laboratory animals. The proof of principle work included the selection of the culture conditions for the preparation of sulphur labelled yeast, the study of the suitability of this labelled yeast as sulphur source for tracer studies using in vitro gastrointestinal digestion and the administration of the 34S-labelled yeast to laboratory animals to follow the fate and distribution of 34S in the organism. For in vitro gastrointestinal digestion, the combination of sodium dodecyl sulphate-polyacrylamide gel electrophoresis and high-performance liquid chromatography and inductively coupled plasma mass spectrometry (HPLC-ICP-MS) showed that labelled methionine, cysteine and other low molecular weight sulphur-containing biomolecules were the major components in the digested extracts of the labelled yeast. Next, in vivo kinetic experiments were performed in healthy Wistar rats after the oral administration of 34S-labelled yeast. The isotopic composition of total sulphur in tissues, urine and faeces was measured by double-focusing inductively coupled plasma mass spectrometry after microwave digestion. It was observed that measurable isotopic enrichments were detected in all samples. Finally, initial investigations on sulphur isotopic composition of serum and urine samples by HPLC-ICP-MS have been carried out. For serum samples, no conclusive data were obtained. Interestingly, chromatographic analysis of urine samples showed differential isotope enrichment for several sulphur-containing biomolecules.

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  1. Villar-Garea A, Griese M, Imhof A (2007) Biomarker discovery from body fluids using mass spectrometry. J Chromatogr B 849:105–114

    Article  CAS  Google Scholar 

  2. Apweiler R, Aslanidis C, Deufel T, Gerstner A, Hansen J, Hochstrasser D et al (2009) Approaching clinical proteomics: current state and future fields of application in fluid proteomics. Clin Chem Lab Med 47:724–744

    Article  CAS  Google Scholar 

  3. Silberring J, Ciborowski P (2010) Biomarker discovery and clinical proteomics. TRAC Trends Anal Chem 29:128–140

    Article  CAS  Google Scholar 

  4. Srinivas PR, Srivastava S, Hanash S, Wright GL (2001) Proteomics in early detection of cancer. Clin Chem 47:1901–1911

    CAS  Google Scholar 

  5. Shau H, Chandler GS, Whitelegge JP, Gornbein JA, Faull KF, Chang HR (2003) Proteomic profiling of cancer biomarkers. Brief Funct Genom Proteomics 2:147–158

    Article  CAS  Google Scholar 

  6. Fliser D, Novak J, Thongboonkerd V, Argilés A, Jankowski V, Girolami MA, Jankowski J, Mischak H (2007) Advances in urinary proteome analysis and biomarker discovery. J Am Soc Nephrol 18:1057–1071

    Article  CAS  Google Scholar 

  7. Ru QC, Katenhusen RA, Zhu LA, Silberman J, Yang S, Orchard TJ, Brzeski H, Liebman M, Ellsworth DL (2006) Proteomic profiling of human urine using multi-dimensional protein identification technology. J Chromatogr A 1111:166–174

    Article  CAS  Google Scholar 

  8. Wittke S, Fliser D, Haubitz M, Bartel S, Krebs R, Hausadel F, Hillmann M, Golovko I, Koester P, Haller H, Kaiser T, Mischak H, Weissinger EM (2003) Determination of peptides and proteins in human urine with capillary electrophoresis–mass spectrometry, a suitable tool for the establishment of new diagnostic markers. J Chromatogr A 1013:173–181

    Article  CAS  Google Scholar 

  9. Haubitz M, Wittke S, Weissinger EM, Walden M, Rupprecht HD, Floege J, Haller H, Mischak H (2005) Urine protein patterns can serve as diagnostic tools in patients with IgA nephropathy. Kidney Int 67:2313–2320

    Article  CAS  Google Scholar 

  10. Theodorescu D, Wittke S, Ross MM, Walden M, Conaway M, Just I, Mischak H, Frierson HF (2006) Discovery and validation of new protein biomarkers for urothelial cancer: a prospective analysis. Lancet Oncol 7:230–240

    Article  CAS  Google Scholar 

  11. Kaiser T, Kamal H, Rank A, Kolb HJ, Holler E, Ganser A, Hertenstein B, Mischak H, Weissinger EM (2004) Proteomics applied to the clinical follow-up of patients after allogeneic hematopoietic stem cell transplantation. Blood 104:340–349

    Article  CAS  Google Scholar 

  12. Wind M, Wegener A, Eisenmenger A, Kellner R, Lehmann WD (2003) Sulfur as the key element for quantitative protein analysis by capillary liquid chromatography coupled to element mass spectrometry. Angew Chem Int Ed 42:3425–3427

    Article  CAS  Google Scholar 

  13. Prange A, Profrock D (2008) Chemical labels and natural element tags for the quantitative analysis of bio-molecules. J Anal At Spectrom 23:432–459

    Article  CAS  Google Scholar 

  14. Rappel C, Schaumlöffel D (2008) The role of sulfur and sulfur isotope dilution analysis in quantitative protein analysis. Anal Bioanal Chem 390:605–615

    Article  CAS  Google Scholar 

  15. Anderson JW (1980) Assimilation of inorganic sulfate into cysteine. In: Stumpf PK, Conn EE (eds) The biochemistry of plants, vol 5. Academic, New York, p 203

    Google Scholar 

  16. Porro D, Sauer M, Branduardi P, Mattanovich D (2005) Recombinant protein production in yeasts. Mol Biotechnol 31:245–259

    Article  CAS  Google Scholar 

  17. Thomas D, Surdin-Kerjan Y (1997) Metabolism of sulfur amino acids in Saccharomyces cerevisiae. Microbiol Mol Biol Rev 61:503–532

    CAS  Google Scholar 

  18. Barnett JA (2003) Beginnings of microbiology and biochemistry: the contribution of yeast research. Microbiology 149:557–567

    Article  CAS  Google Scholar 

  19. Giner Martínez-Sierra J, Moreno Sanz F, Herrero Espílez P, Marchante Gayón JM, García Alonso JI (2007) Biosynthesis of sulfur-34 labelled yeast and its characterisation by multicollector-ICP-MS. J Anal At Spectrom 22:1105–1112

    Article  Google Scholar 

  20. Rodríguez-González P, Ruiz Encinar J, García Alonso JI, Sanz-Medel A (2005) Monitoring the degradation and solubilisation of butyltin compounds during in vitro gastrointestinal digestion using “triple spike” isotope dilution GC-ICP-MS. Anal Bioanal Chem 381:380–387

    Article  Google Scholar 

  21. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Article  CAS  Google Scholar 

  22. Rodríguez-González P, García Alonso JI (2010) Recent advances in isotope dilution analysis for elemental speciation. J Anal At Spectrom 25:239–259

    Article  Google Scholar 

  23. Giner Martínez-Sierra J, Moreno Sanz F, Herrero Espílez P, Santamaria-Fernandez R, Marchante Gayón JM, García Alonso JI (2010) Evaluation of different analytical strategies for the quantification of sulfur-containing biomolecules by HPLC-ICP-MS: application to the characterisation of 34S-labelled yeast. J Anal At Spectrom 25:989–997

    Article  Google Scholar 

  24. Rodríguez-González P, Marchante-Gayón JM, García Alonso JI, Sanz-Medel A (2005) Isotope dilution analysis for elemental speciation: a tutorial review. Spectrochim Acta B 60:151–207

    Article  Google Scholar 

  25. Giner Martínez-Sierra J, Santamaria-Fernandez R, Hearn R, Marchante Gayón JM, García Alonso JI (2010) Development of a direct procedure for the measurement of sulfur isotope variability in beers by MC-ICP-MS. J Agric Food Chem 58:4043–4050

    Article  Google Scholar 

  26. Santamaria-Fernandez R, Giner Martínez-Sierra J, Marchante Gayón JM, García Alonso JI, Hearn R (2009) Measurement of longitudinal sulfur isotopic variations by laser ablation MC-ICP-MS in single human hair strands. Anal Bioanal Chem 394:225–233

    Article  CAS  Google Scholar 

  27. Tirumalai RS, Chan KC, Prieto DA, Issaq HJ, Conrads TP, Veenstra TD (2003) Characterisation of the low molecular weight human serum proteome. Mol Cell Proteomics 2:1096–1103

    Article  CAS  Google Scholar 

  28. Shou M, Conrads TP, Veenstra TD (2005) Proteomic approaches to biomarker detection. Brief Funct Genom Proteomics 4:69–75

    Article  Google Scholar 

  29. Luque-Garcia JL, Neubert TA (2007) Sample preparation for serum/plasma profiling and biomarker identification by mass spectrometry. J Chromatogr A 1153:259–276

    Article  CAS  Google Scholar 

  30. Martorella A, Robbins R (2007) Serum peptide profiling: identifying novel cancer biomarkers for early disease detection. Acta Biomed 78:123–128

    Google Scholar 

  31. Linke T, Doraiswamy S, Harrison EH (2007) Rat plasma proteomics: effects of abundant protein depletion on proteomic analysis. J Chromatogr B 849:273–281

    Article  CAS  Google Scholar 

  32. Khan A, Packer NH (2006) Simple urinary sample preparation for proteomic analysis. J Proteome Res 5:2824–2838

    Article  CAS  Google Scholar 

  33. Magagnotti C, Fermo I, Carletti RM, Ferrari M, Bachi A (2010) Comparison of different depletion strategies for improving resolution of the human urine proteome. Clin Chem Lab Med 48:531–535

    Article  CAS  Google Scholar 

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This work was supported by the Ministry of Science and Innovation, Madrid, Spain (project CTQ2009-12814) and the Education and Science Council of the Principado de Asturias (grant BP07-059). Teresa Fernández and Agustín Brea from the Biotery of the University of Oviedo are gratefully acknowledged for their help and kind suggestions. The authors thank Rafael Peláez for his assistance in the experimental work regarding gels.

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Correspondence to J. I. García Alonso.

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Published in the topical collection Isotope Ratio Measurements: New Developments and Applications with guest editors Klaus G. Heumann and Torsten C. Schmidt.

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Martínez-Sierra, J.G., Sanz, F.M., Espílez, P.H. et al. Sulphur tracer experiments in laboratory animals using 34S-labelled yeast. Anal Bioanal Chem 405, 2889–2899 (2013).

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