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

Jasna Nikolić, Andrijana Nešić, Milena Čavić, Neda Đorđević, Uroš Anđelković, Marina Atanasković-Marković, Branko Drakulić, Marija Gavrović-Jankulović

Journal: Biochimica et biophysica acta. General subjects 2017;1861(2):126-134

PMID: 27864150

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Abstract

BACKGROUND

Protein oxidation can occur as a consequence of lipid peroxidation during food processing. The aim of this work was to explore the 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 modification in 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 expression for pro-inflammatory cytokines and protein secretion.

RESULTS

Out of 9 predicted, five modified Lys residues were confirmed by MS/MS analysis: TQINKVVR, DILNQITKPNDVYSFSLASR, YPILPEYLQCVKELYR, AFKDEDTQAMPFR, KIKVYLPR, and IKVYLPR. The introduced MDA modifications influenced profile of IgE reactivity to OVA. Treatment of T84 epithelial cells with OVA and OVA modified with 1mM MDA, induced up-regulation of pro-inflammatory cytokines (IL-1β, IL-25, IL-33, TSLP and TNFα), while OVA modification with 10mM MDA induced down regulation of the cytokine expression profile, except for IL-1β. OVA and OVA modified with 1mM MDA induced secretion of epithelial cells specific cytokine IL-33.

CONCLUSIONS

This finding indicated that OVA and its MDA modified form have the potential to trigger the innate immunity by inducing up-regulation and secretion of pro-allergenic IL-33 in T84 intestinal epithelial cells.

GENERAL SIGNIFICANCE

Interactions of ovalbumin and its MDA modified form with intestinal epithelial cells can induce a specific immunological priming necessary for the downstream activation of innate immunity.

Address: Department of Biochemistry, Faculty of Chemistry, University of Belgrade, Belgrade, Serbia.; Department of Experimental Oncology, Institute for Oncology and Radiology of Serbia, Belgrade, Serbia.; Centre for High-Throughput Technologies, Department of Biotechnology, University of Rijeka, Rijeka, Croatia; Department of Chemistry, Institute of Chemistry, Technology and Metallurgy, University of Belgrade, Belgrade, Serbia.; Medical Faculty, University of Belgrade, Belgrade, Serbia; University Children's Hospital of Belgrade, Belgrade, Serbia.; Department of Chemistry, Institute of Chemistry, Technology and Metallurgy, University of Belgrade, Belgrade, Serbia.; Department of Biochemistry, Faculty of Chemistry, University of Belgrade, Belgrade, Serbia. Electronic address: [email protected].

DOI: 10.1016/j.bbagen.2016.11.021

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MeSH Terms: Cell Line, Cytokines, Epithelial Cells, Humans, Immunity, Innate, Immunoglobulin E, Inflammation, Interleukin-1beta, Interleukin-33, Intestinal Mucosa, Intestines, Malondialdehyde, Molecular Dynamics Simulation, Ovalbumin, Tandem Mass Spectrometry, Up-Regulation, Thymic Stromal Lymphopoietin