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Mass spectrometry of in-gel digests reveals differences in amino acid sequences of high-molecular-weight glutenin subunits in spelt and emmer compared to common wheat

  • Sabrina Geisslitz
  • Antoine H. P. America
  • Katharina Anne ScherfEmail author
Research Paper

Abstract

High-molecular-weight glutenin subunits (HMW-GS) play an important role for the baking quality of wheat. The ancient wheats emmer and spelt differ in their HMW-GS pattern compared to modern common wheat and this might be one reason for their comparatively poor baking quality. The aim of this study was to elucidate similarities and differences in the amino acid sequences of two 1Bx HMW-GS of common wheat, spelt and emmer. First, the sodium dodecyl polyacrylamide gel electrophoresis (SDS-PAGE) system was optimized to separate common wheat, spelt and emmer Bx6 and Bx7 from other HMW-GS (e.g., 1Ax and 1By) in high concentrations. The in-gel digests of the Bx6 and Bx7 bands were analyzed by untargeted LC-MS/MS experiments revealing different UniProtKB accessions in spelt and emmer compared to common wheat. The HMW-GS Bx6 and Bx7, respectively, of emmer and spelt showed differences in the amino acid sequences compared to those of common wheat. The identities of the peptide variations were confirmed by targeted LC-MS/MS. These peptides can be used to differentiate between Bx6 and Bx7 of spelt and emmer and Bx6 and Bx7 of common wheat. The findings should help to increase the reliability and curation status of wheat protein databases and to understand the effects of protein structure on the functional properties.

Graphical abstract

Keywords

High-molecular-weight glutenin subunits (HMW-GS) LC-MS/MS Proteomics SDS-PAGE Wheat 

Notes

Acknowledgments

The authors thank Sami Kaviani-Nejad and Katharina Booz (Leibniz-LSB@TUM) and Bert Schipper (Wageningen UR) for excellent technical assistance.

Author contributions

All authors contributed to the study conception and design. Experimental work, data analysis, and preparation of tables and figures were performed by Sabrina Geisslitz. Data analysis was additionally done by Antoine H. P. America. The first draft of the manuscript was written by Sabrina Geisslitz and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

Funding information

This IGF Project of the FEI was supported via AiF within the programme for promoting the Industrial Collective Research (IGF) of the German Ministry of Economic Affairs and Energy (BMWi), based on a resolution of the German Parliament. Project AiF 18355 N. This submission was supported by the Fachgruppe Analytische Chemie of the Gesellschaft Deutscher Chemiker via a scholarship for finalizing a publication in a cooperation project. Additional funding came from the Technical University of Munich (TUM) through the TUM Graduate School (GS) Internationalization Grant.

Compliance with ethical standards

No human or animal subjects were used in the study. No informed consent was required for this study.

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

216_2019_2341_MOESM1_ESM.pdf (1.1 mb)
ESM 1 (PDF 1129 kb)

References

  1. 1.
    Goesaert H, Brijs K, Veraverbeke WS, Courtin CM, Gebruers K, Delcour J. A. Wheat flour constituents: how they impact bread quality, and how to impact their functionality. Trends Food Sci Technol. 2005;16:12–30.CrossRefGoogle Scholar
  2. 2.
    Osborne TB. The proteins of the wheat kernel. Washington: Carnegie Institution; 1907.CrossRefGoogle Scholar
  3. 3.
    Wieser H. Chemistry of gluten proteins. Food Microbiol. 2007;24(2):115–9.PubMedCrossRefPubMedCentralGoogle Scholar
  4. 4.
    Payne PI. Genetics of wheat storage proteins and the effect of allelic variation on breadmaking quality. Annu Rev Plant Physiol. 1987;38:141–53.CrossRefGoogle Scholar
  5. 5.
    Payne PI, Corfield G, Holt LM, Blackman JA. Correlation between the inheritance of certain high molecular weight subunits of glutenin and bread making quality in progenies of six crosses of bread wheat. J Sci Food Agric. 1981;32:51–60.CrossRefGoogle Scholar
  6. 6.
    Payne PI, Law CN, Mudd EE. Control by homologous group 1 chromosomes of the high-molecular-weight subunits of glutenin, a major protein of wheat endosperm. Theor Appl Genet. 1980;58:113–20.PubMedCrossRefPubMedCentralGoogle Scholar
  7. 7.
    Payne PI, Lawrence GJ. Catalogue of alleles for the complex gene loci Glu-A1, Glu-B1 and Glu-D1 which code for high-molecular weight subunits of glutenin in hexaploid wheat. Cereal Res Commun. 1983;11:29–35.Google Scholar
  8. 8.
    Payne PI, Holt LM, Law CN. Structural and genetical studies on the high-molecular-weight subunits of wheat glutenin. Theor Appl Genet. 1981;60:229–36.PubMedCrossRefPubMedCentralGoogle Scholar
  9. 9.
    Jiang P, Xue J, Duan L, Gu Y, Mu J, Han S, et al. Effects of high-molecular-weight glutenin subunit combination in common wheat on the quality of crumb structure. J Sci Food Agric. 2019;99(4):1501–8.PubMedCrossRefPubMedCentralGoogle Scholar
  10. 10.
    Shewry PR, Halford NG, Tatham AS. High molecular weight subunits of wheat glutenin. J Cereal Sci. 1992;15(2):105–20.CrossRefGoogle Scholar
  11. 11.
    Geisslitz S, Wieser H, Scherf KA, Koehler P. Gluten protein composition and aggregation properties as predictors for bread volume of common wheat, spelt, durum wheat, emmer and einkorn. J Cereal Sci. 2018;83:204–12.CrossRefGoogle Scholar
  12. 12.
    Schalk K, Lexhaller B, Koehler P, Scherf KA. Isolation and characterization of gluten protein types from wheat, rye, barley and oats for use as reference materials. PLoS One. 2017;12(2):e0172819 1981;32:359–71.PubMedPubMedCentralCrossRefGoogle Scholar
  13. 13.
    Wieser H, Antes S, Seilmeier W. Quantitative determination of gluten protein types in wheat flour by reversed-phase high-performance liquid chromatography. Cereal Chem. 1998;75(5):644–50.CrossRefGoogle Scholar
  14. 14.
    Dong K, Hao C, Wang A, Cai M, Yan Y. Characterization of HMW glutenin subunits in bread and tetraploid wheats by reversed-phase high-performance liquid chromatography. Cereal Res Commun. 2009;37(1):65–73.CrossRefGoogle Scholar
  15. 15.
    Qian Y, Preston K, Krokhin O, Mellish J, Ens W. Characterization of wheat gluten proteins by HPLC and MALDI TOF mass spectrometry. J Am Soc Mass Spectrom. 2008;19(10):1542–50.PubMedCrossRefPubMedCentralGoogle Scholar
  16. 16.
    Marchylo BA, Kruger JE, Hatcher DW. Quantitative reversed-phase high-performance liquid chromatographic analysis of wheat storage proteins as a potential quality prediction tool. J Cereal Sci. 1989;9(2):113–30.CrossRefGoogle Scholar
  17. 17.
    Lagrain B, Brunnbauer M, Rombouts I, Koehler P. Identification of intact high molecular weight glutenin subunits from the wheat proteome using combined liquid chromatography-electrospray ionization mass spectrometry. PLoS One. 2013;8(3):e58682.PubMedPubMedCentralCrossRefGoogle Scholar
  18. 18.
    Gao L, Ma W, Chen J, Wang KE, Li J, Wang S, et al. Characterization and comparative analysis of wheat high molecular weight glutenin subunits by SDS-PAGE, RP-HPLC, HPCE, and MALDI-TOF-MS. J Agric Food Chem. 2010;58(5):2777–86.PubMedCrossRefPubMedCentralGoogle Scholar
  19. 19.
    Zhang Q, Dong Y, An X, Wang A, Zhang Y, Li X, et al. Characterization of HMW glutenin subunits in common wheat and related species by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). J Cereal Sci. 2008;47(2):252–61.CrossRefGoogle Scholar
  20. 20.
    Liu L, Wang A, Appels R, Ma J, Xia X, Lan P, et al. A MALDI-TOF based analysis of high molecular weight glutenin subunits for wheat breeding. J Cereal Sci. 2009;50(2):295–301.CrossRefGoogle Scholar
  21. 21.
    Jin M, Xie ZZ, Ge P, Li J, Jiang SS, Subburaj S, et al. Identification and molecular characterisation of HMW glutenin subunit 1By16* in wild emmer. J Appl Genet. 2012;53(3):249–58.PubMedCrossRefPubMedCentralGoogle Scholar
  22. 22.
    Barro F, Rooke L, Békés F, Gras P, Tatham AS, Fido R, et al. Transformation of wheat with high molecular weight subunit genes results in improved functional properties. Nat Biotechnol. 1997;15(12):1295–9.PubMedCrossRefPubMedCentralGoogle Scholar
  23. 23.
    Blatter RHE, Jacomet S, Schlumbaum A. About the origin of European spelt (Triticum spelta L.): allelic differentiation of the HMW Glutenin B1-1 and A1-2 subunit genes. Theor Appl Genet. 2004;108:360.PubMedCrossRefPubMedCentralGoogle Scholar
  24. 24.
    Dupont FM, Vensel WH, Tanaka CK, Hurkman WJ, Altenbach SB. Deciphering the complexities of the wheat flour proteome using quantitative two-dimensional electrophoresis, three proteases and tandem mass spectrometry. Proteome Sci. 2011;9:10.PubMedPubMedCentralCrossRefGoogle Scholar
  25. 25.
    Colgrave ML, Byrne K, Howitt CA. Food for thought: selecting the right enzyme for the digestion of gluten. Food Chem. 2017;234:389–97.PubMedCrossRefPubMedCentralGoogle Scholar
  26. 26.
    Oezbek O, Goeçmen Taşkin B, Keskin Şan S, Eser V, Arslan O. High-molecular-weight glutenin subunit variation in Turkish emmer wheat [Triticum turgidum L. ssp. dicoccon (Schrank) Thell.] landraces. Plant Syst Evol. 2012;298:1795–804.CrossRefGoogle Scholar
  27. 27.
    Pflueger LA, Martín LM, Alvarez JB. Variation in the HMW and LMW glutenin subunits from Spanish accessions of emmer wheat (Triticum turgidum ssp. Dicoccum Schrank). Theor Appl Genet. 2001;102:767–72.CrossRefGoogle Scholar
  28. 28.
    Kasarda DD, Woodard KM, Adalsteins AE. Resolution of high molecular weight glutenin subunits by a new SDS-PAGE system incorporating a neutral pH buffer. Cereal Chem. 1998;75:70e71.CrossRefGoogle Scholar
  29. 29.
    Lagrain B, Rombouts I, Wieser H, Delcour JA, Koehler P. A reassessment of the electrophoretic mobility of high molecular weight glutenin subunits of wheat. J Cereal Sci. 2012;56:726–32.CrossRefGoogle Scholar
  30. 30.
    Shevchenko A, Tomas H, Havlis J, Olsen JV, Mann M. In-gel digestion for mass spectrometric characterization of proteins and proteomes. Nat Protoc. 2006;1(6):2856–60.PubMedCrossRefPubMedCentralGoogle Scholar
  31. 31.
    Cox J, Neuhauser N, Michalski A, Scheltema RA, Olsen JV, Mann M. Andromeda: a peptide search engine integrated into the MaxQuant environment. J Proteome Res. 2011;10:1794–805.PubMedCrossRefPubMedCentralGoogle Scholar
  32. 32.
    Ma B, Zhang K, Hendrie C, Liang C, Li M, Doherty-Kirby A, et al. PEAKS: powerful software for peptide de novo sequencing by tandem mass spectrometry. Rapid Commun Mass Spectrom. 2003;17(20):2337–42.PubMedCrossRefPubMedCentralGoogle Scholar
  33. 33.
    MacLean B, Tomazela DM, Shulman N, Chambers M, Finney GL, Frewen B, et al. Skyline: an open source document editor for creating and analyzing targeted proteomics experiments. Bioinformatics. 2010;26:966–8.PubMedPubMedCentralCrossRefGoogle Scholar
  34. 34.
    Gessendorfer B, Koehler P, Wieser H. Preparation and characterization of enzymatically hydrolyzed prolamins from wheat, rye, and barley as references for the immunochemical quantitation of partially hydrolyzed gluten. Anal Bioanal Chem. 2009;395(6):1721–8.PubMedCrossRefPubMedCentralGoogle Scholar
  35. 35.
    Scarnato L, Gadermaier G, Volta U, De Giorgio R, Caio G, Lanciotti R, et al. Immunoreactivity of gluten-sensitized sera toward wheat, rice, corn, and amaranth flour proteins treated with microbial transglutaminase. Front Microbiol. 2019;10:470.Google Scholar
  36. 36.
    Wieser H, Scherf KA. Preparation of a defined gluten hydrolysate for diagnosis and clinical investigations of wheat hypersensitivities. Nutrients. 2018;10:1411.PubMedCentralCrossRefGoogle Scholar
  37. 37.
    Wan Y, Gritsch CS, Hawkesford MJ, Shewry PR. Effects of nitrogen nutrition on the synthesis and deposition of the omega-gliadins of wheat. Ann Bot. 2014;113(4):607–15.PubMedCrossRefPubMedCentralGoogle Scholar
  38. 38.
    Pastorello EA, Farioli L, Conti A, Pravettoni V, Bonomi S, Iametti S, et al. Wheat IgE-mediated food allergy in European patients: α-amylase inhibitors, lipid transfer proteins and low-molecular-weight glutenins. Int Arch Allergy Immunol. 2007;144(1):10–22.PubMedCrossRefPubMedCentralGoogle Scholar
  39. 39.
    Marchetti-Deschmann M, Lehner A, Peterseil V, Soevegjarto F, Hochegger R, Allmaier G. Fast wheat variety classification by capillary gel electrophoresis-on-a-chip after single-step one-grain high molecular weight glutenin extraction. Anal Bioanal Chem. 2011;400:2403–14.PubMedCrossRefPubMedCentralGoogle Scholar
  40. 40.
    Ludwig C, Aebersold R. Getting absolute: determining absolute protein quantities via selected reaction monitoring mass spectrometry. In: Eyers CE, Gaskell S, editors. Quantitative Proteomics. London: The Royal Society of Chemistry; 2014. pp. 80–109.Google Scholar
  41. 41.
    Bogdanow B, Zauber H, Selbach M. Systematic errors in peptide and protein identification and quantification by modified peptides. Mol Cell Proteomics. 2016;15:2791–801.PubMedPubMedCentralCrossRefGoogle Scholar
  42. 42.
    Rechenberger J, Samaras P, Jarzab A, Behr J, Frejno M, Djukovic A, et al. Challenges in clinical metaproteomics highlighted by the analysis of acute leukemia patients with gut colonization by multidrug-resistant Enterobacteriaceae. Proteomes. 2019;7:2.PubMedCentralCrossRefGoogle Scholar
  43. 43.
    The International Wheat Genome Sequencing Consortium (IWGSC). Shifting the limits in wheat research and breeding using a fully annotated reference genome. Science. 2018;361:eaar7191.PubMedCrossRefPubMedCentralGoogle Scholar
  44. 44.
    Payne PI, Nightingale MA, Krattiger AF, Holt LM. The relationship between HMW glutenin subunit composition and the bread-making quality of British-grown wheat varieties. J Sci Food Agric. 1987;40:51–65.CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Leibniz-Institute for Food Systems Biology at the Technical University of MunichFreisingGermany
  2. 2.Wageningen Plant ResearchWageningen University & ResearchWageningenThe Netherlands
  3. 3.Department of Bioactive and Functional Food ChemistryInstitute of Applied Biosciences, Karlsruhe Institute of Technology (KIT)KarlsruheGermany

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