Proteomic analysis of wheat α/A- and β-gliadins

Abstract

Gliadins from the Polish common winter wheat cultivar Sukces were analysed by analytical and preparative A-PAGE combined with 2-DE method. The main aim of this study was to identify the highest possible number of a/A- and β-gliadin fractions. Gliadins from the wheat cv. Sukces were separated by 2-DE into 82 spots. Preparative A-PAGE combined with the 2-DE method supported the identification of the analysed gliadin fractions. 12 spots were identified as typical a/A-gliadins out of 40 and 7 as typical β-gliadins out of 15 separated by 2-DE.  

Low-temperature crosslinking of proteins using non-toxic citric acid in neutral aqueous medium: Mechanism and kinetic study

“…However, without adding alkaline, citric acid decreased the pH of the dispersion to less than 3, much lower than the isoelectric point of gliadin (pH 6.5) (Dziuba et al, 2014). Therefore, overwhelmingly more amine groups might carry positive charge, and thus would not attack carbonyl groups to initiate the nucleophilic substitution.…”

CRISPR/Cas9 Gene Editing of Gluten in Wheat to Reduce Gluten Content and Exposure—Reviewing Methods to Screen for Coeliac Safety

“…Acid-PAGE or SDS-PAGE has been used in all RNAi and CRISPR/Cas9 studies of gliadin genes published to date ( Table 1). However, one-dimensional PAGE does not give full information on gluten proteins, since the protein spots overlap, as is visible in 2D-electrophoresis (68)(69)(70)(71)(72)(73). Changes in the Acid-PAGE or SDS-PAGE one-dimensional gel electrophoresis profiles should therefore be confirmed by proteomic techniques.…”

Comparison of MALDI-TOF-MS and RP-HPLC as Rapid Screening Methods for Wheat Lines With Altered Gliadin Compositions

“…For MALDI-TOF-MS analysis, gliadin was prepared as described by Dziuba et al (2014). Thirty mg of flour was mixed with 150 µl 0.15 M NaCl solution and shaken for 2 h. After centrifugation at 12,000 rpm for 10 min, the supernatant containing albumin/globulin was discarded.…”

“…To verify the effects of extraction solvents in mass spectra, we optimized the gliadin protein extraction from whole wheat grains according to the two previously reported methods (Dziuba et al, 2014;Han et al, 2015). The difference between the two methods is the use of 0.15 M NaCl to remove salt-soluble albumins and globulins before extracting gliadins with aqueous alcohol.…”