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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.
Anal Bioanal Chem. 2020 Feb; 412(6):1277-1289.AB

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.

Authors+Show Affiliations

Leibniz-Institute for Food Systems Biology at the Technical University of Munich, 85354, Freising, Germany.Wageningen Plant Research, Wageningen University & Research, 6708 PB, Wageningen, The Netherlands.Leibniz-Institute for Food Systems Biology at the Technical University of Munich, 85354, Freising, Germany. katharina.scherf@kit.edu. Department of Bioactive and Functional Food Chemistry, Institute of Applied Biosciences, Karlsruhe Institute of Technology (KIT), 76131, Karlsruhe, Germany. katharina.scherf@kit.edu.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

31927602

Citation

Geisslitz, Sabrina, et al. "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." Analytical and Bioanalytical Chemistry, vol. 412, no. 6, 2020, pp. 1277-1289.
Geisslitz S, America AHP, Scherf KA. 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. Anal Bioanal Chem. 2020;412(6):1277-1289.
Geisslitz, S., America, A. H. P., & Scherf, K. A. (2020). 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. Analytical and Bioanalytical Chemistry, 412(6), 1277-1289. https://doi.org/10.1007/s00216-019-02341-9
Geisslitz S, America AHP, Scherf KA. 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. Anal Bioanal Chem. 2020;412(6):1277-1289. PubMed PMID: 31927602.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - 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. AU - Geisslitz,Sabrina, AU - America,Antoine H P, AU - Scherf,Katharina Anne, Y1 - 2020/01/11/ PY - 2019/10/17/received PY - 2019/12/09/accepted PY - 2019/11/26/revised PY - 2020/1/14/pubmed PY - 2020/4/3/medline PY - 2020/1/14/entrez KW - High-molecular-weight glutenin subunits (HMW-GS) KW - LC-MS/MS KW - Proteomics KW - SDS-PAGE KW - Wheat SP - 1277 EP - 1289 JF - Analytical and bioanalytical chemistry JO - Anal Bioanal Chem VL - 412 IS - 6 N2 - 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. SN - 1618-2650 UR - https://www.unboundmedicine.com/medline/citation/31927602/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_ L2 - https://dx.doi.org/10.1007/s00216-019-02341-9 DB - PRIME DP - Unbound Medicine ER -