Effect of malondialdehyde on the ovalbumin structure and its interactions with T84 epithelial cells

Nikolić Jasna; Nešić Andrijana; Čavić Milena; Đorđević Neda; Anđelković Uroš; Atanacković-Marković M; Drakulić Branko; Gavrović-Jankulović Marija

doi:10.1016/j.bbagen.2016.11.021

Published November 14, 2016 | Version v.1

Journal article Open

Effect of malondialdehyde on the ovalbumin structure and its interactions with T84 epithelial cells

1. Faculty of Chemistry, University of Belgrade, Belgrade, Serbia
2. Department of Experimental Oncology, Institute for Oncology and Radiology of Serbia, Belgrade, Serbia
3. Centre for High-Throughput Technologies, Department of Biotechnology, University of Rijeka, Rijeka, Croatia and Department of Chemistry, Institute of Chemistry, Technology and Metallurgy, University of Belgrade, Belgrade, Serbia
4. Medical Faculty, University of Belgrade, Serbia and University Children's Hospital of Belgrade, Serbia
5. Department of Chemistry, Institute of Chemistry, Technology and Metallurgy, University of Belgrade, Belgrade, Serbia
6. Faculty of Chemistry, University of Belgrade, Belgrade, Serbia and University Children's Hospital of Belgrade, Belgrade, Serbia

Background: Protein oxidation can occur as a consequence of lipid peroxidation during food processing. The aim of this work was to explorethe effect of malondialdehyde (MDA) modification of ovalbumin (OVA) on its interaction with T84 intestinal cells. Methods: Molecular dynamics simulation was employed for the prediction of MDA modificationin the OVA, while introduced structural changes were evaluated by measurement of carbonyl group content, fluorescence spectra,MS/MS analysis, and IgE reactivity. Effects of MDA modified OVA on T84 epithelial cells were analyzed by gene expressionfor pro-inflammatorycytokines andprotein secretion.Results: Out of9 predicted,five modified Lys residues were confirmed by MS/MS analysis:51TQINKVVR58, 85DILNQITKPNDVYSFSLASR104, 111YPILPEYLQCVKELYR126, 187AFKDEDTQAMPFR199, 277KIKVYLPR284, and 278IKVYLPR284. The introduced MDA modificationsinfluencedprofileof IgE reactivity toOVA. Treatment of T84 epithelial cells with OVA and OVA modified with 1 mM MDA,induced up-regulationof pro-inflammatorycytokines (IL-1β, IL-25, IL-33, TSLP and TNFα), while OVA modification with 10 mM MDAinduceddown regulationof the cytokineexpressionprofile,except for IL-1β. OVA and OVA modified with 1 mM MDA inducedsecretion of epithelial cells specific cytokine IL-33. Conclusions: This finding indicated that OVAand its MDA modifiedformhavethe potential to trigger the innate immunity by inducing up-regulationand secretion of pro-allergenic IL-33in T84 intestinal epithelial cells.General significance:Interactions ofovalbumin and its MDA modified form with intestinal epithelial cells can induce a specific immunological priming necessary for the downstream activation of innate immunity.

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