-
Tytuł:
-
Hybrid QconCAT-Based Targeted Absolute and Data-Independent Acquisition-Based Label-Free Quantification Enables In-Depth Proteomic Characterization of Wheat Amylase/Trypsin Inhibitor Extracts.
-
Autorzy:
-
Sielaff M; Institute for Immunology, University Medical Center of the Johannes Gutenberg-University Mainz, Langenbeckstr. 1, 55131 Mainz, Germany.
Curella V; Institute of Translational Immunology, University Medical Center of the Johannes Gutenberg-University Mainz, Langenbeckstr. 1, 55131 Mainz, Germany.
Neerukonda M; Institute of Translational Immunology, University Medical Center of the Johannes Gutenberg-University Mainz, Langenbeckstr. 1, 55131 Mainz, Germany.
Afzal M; State Plant Breeding Institute, University of Hohenheim, Fruwirthstr. 21, 70599 Stuttgart, Germany.
El Hassouni K; State Plant Breeding Institute, University of Hohenheim, Fruwirthstr. 21, 70599 Stuttgart, Germany.
Distler U; Institute for Immunology, University Medical Center of the Johannes Gutenberg-University Mainz, Langenbeckstr. 1, 55131 Mainz, Germany.
Schuppan D; Institute of Translational Immunology, University Medical Center of the Johannes Gutenberg-University Mainz, Langenbeckstr. 1, 55131 Mainz, Germany.; Division of Gastroenterology, Beth Israel Deaconess Medical Center, Harvard Medical School, 330 Brookline Avenue, Boston, Massachusetts 02215, United States.
Longin CFH; State Plant Breeding Institute, University of Hohenheim, Fruwirthstr. 21, 70599 Stuttgart, Germany.
Tenzer S; Institute for Immunology, University Medical Center of the Johannes Gutenberg-University Mainz, Langenbeckstr. 1, 55131 Mainz, Germany.
-
Źródło:
-
Journal of proteome research [J Proteome Res] 2021 Mar 05; Vol. 20 (3), pp. 1544-1557. Date of Electronic Publication: 2021 Jan 28.
-
Typ publikacji:
-
Journal Article; Research Support, Non-U.S. Gov't
-
Język:
-
English
-
Imprint Name(s):
-
Original Publication: Washington, D.C. : American Chemical Society, c2002-
-
MeSH Terms:
-
Triticum*/genetics
Trypsin Inhibitors*
Amylases ; Plant Breeding ; Plant Extracts ; Proteomics ; Reproducibility of Results ; Trypsin
-
Contributed Indexing:
-
Keywords: QconCAT; Triticum aestivum; alpha-amylase/trypsin inhibitor; bottom-up proteomics; celiac disease; data-independent acquisition; flour; label-free quantification; non-celiac wheat sensitivity; wheat
-
Substance Nomenclature:
-
0 (Plant Extracts)
0 (Trypsin Inhibitors)
EC 3.2.1.- (Amylases)
EC 3.4.21.4 (Trypsin)
-
Entry Date(s):
-
Date Created: 20210128 Date Completed: 20210617 Latest Revision: 20210617
-
Update Code:
-
20240105
-
DOI:
-
10.1021/acs.jproteome.0c00752
-
PMID:
-
33507751
-
Wheat amylase/trypsin inhibitors (ATIs) have gained significant relevance as inducers of intestinal and extra-intestinal inflammation. In this study, we present a novel hybrid data-independent acquisition (DIA) liquid chromatography-mass spectrometry (LC-MS) approach, combining QconCAT technology with short microflow LC gradients and DIA and apply the method toward the quantitative proteome analysis of ATI extracts. The presented method is fast, robust, and reproducible and provides precise QconCAT-based absolute quantification of major ATI proteins while simultaneously quantifying the proteome by label-free quantification (LFQ). We analyzed extracts of 60 varieties of common wheat grown in replication and evaluated the reproducibility and precision of the workflow for the quantification of ATIs. Applying the method to analyze different wheat species (i.e., common wheat, spelt, durum wheat, emmer, and einkorn) and comparing the results to published data, we validated inter-laboratory and cross-methodology reproducibility of ATI quantification, which is essential in the context of large-scale breeding projects. Additionally, we applied our workflow to assess environmental effects on ATI expression, analyzing ATI content and proteome of same varieties grown at different locations. Finally, we explored the potential of combining QconCAT-based absolute quantification with DIA-based LFQ proteome analysis for the generation of new hypotheses or assay development.