ENVIRONMENTAL STRESS PROMOTES GLYCATION OF PLANT PROTEINS.

Tatiana Bilova, Elena Lukasheva, Gagan Paudel, Domonik Brauch, Elena Tarakhovskaya, Ahyoung Kim, Thomas Vogt, G.U. Balcke, Claudia Birkemeyer, L.A. Wessjohann, Andrej Frolov

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Abstract

In nature, plants are often subjected to enhanced sun irradiation, heat, weak or moderate drought for relatively long periods of time. The plant response to such environmental stress is manifested with simultaneous enhancement of ROS generation and up-regulation of carbohydrate biosynthesis. This might enhance protein glycation, i.e. interaction of carbonyl compounds (carbohydrates and α-dicarbonyls) with lysyl and arginyl side-chains yielding early and advanced glycation end-products (AGEs). AGEs are known to modulate protein structures and functions, and, therefore, might contribute to stress-related protein damage. They are known to be toxic due to their pro-inflammatory properties, clearly exposed in mammals. Generation of AGEs is well-characterized in animal, but not in plant cells. Recently, accumulation of AGE-modified amino acids was demonstrated in the protein extracts of Arabidopsis thaliana plants subjected to environmental stress [1]. Later, we characterized plant glycation patterns on a proteome level [2]. Recently, we demonstrated, that even weak drought resulted in enhanced glycation at specific protein sites [3]. Here we report the changes in Arabidopsis glycated proteome, induced by high light.
Seven week-old A. thaliana plants, grown under short day (8h light /16h darkness) conditions, were subjected to high light (700 μmol photons m-2 s-1) in parallel to normally irradiated controls (150 μmol photons m-2 s-1). After this period, the treated plants were transferred to the normal light conditions for further three weeks (recovery). The plant stress response was characterized by a panel of biochemical stress markers. Direct and delayed (i.e. observed during the recovery period) effects of high light stress on plant proteome and metabolome were assessed by bottom-up proteomics (LCxLC-ESI-Orbitrap-LIT-MS/MS data-dependent acquisition experiments) and metabolomics (GC-EI-Q-MS).
The high light-treated plants showed signs of oxidative stress, as could be judged from increased levels of H2O2, lipid hydroperoxides and thiobarbiturate (TBA)-reactive compounds. However, the contents of α-dicarbonyl contents and GSH/GSSG ratio were increased, whereas the ascorbate/dehydroascorbate ratio was not changed. During the recovery phase, all mentioned stress markers recovered to the initial levels, while α-dicarbonyls restored their originally high levels. This period was also accompanied with ascorbic acid depletion and pronounced accumulation of carbohydrates. The proteomic analysis revealed only negligible qualitative alterations in Arabidopsis glycated proteome: only 12 and 8 proteins comprising 14 and 8 AGE modifications were found to be specific for light stress and recovery periods, respectively. Moreover, the total number of glycated proteins was not changed under enhanced irradiation and was even lower at the end of the recovery time (641 AGE-modified proteins vs 714 in control). This decrease in the number of glycated proteins could be explained by activation of some enzymatic systems eliminating either AGE precursors, or AGE-modified proteins. Further reactions of AGEs to form new unknown products also can be considered. In contrast, the quantitative stress -related changes in glycated proteome were much more pronounced. Thus, more than 150 AGE modification sites were found to be significantly up-regulated, while the abundance of approximately 50 individual AGE-modified positions was decreased. The glycated polypeptides were mostly involved in transcriptional regulation and protein degradation. Remarkably, the effect of light stress could be observed only after recovery phase, but not directly after restoration of light conditions. It can be explained by high carbohydrate contents in this phase. This assumption is in agreement with a simultaneous increase of dicarbonyl levels that might indicate enhancement of carbohydrate autoxidation. Based on the obtained results we conclude, that light stress causes strong quantitative changes in glycation patterns occurring only at specific sites. Thus, existence of stress-related hotspots of glycation can be assumed. Such glycated proteins accumulate during recovery period but not during the stress application period, that might indicate activation of antiglycative defence under stress conditions.
The research was supported by the Russian Science Foundation (project № 17-16-01042).
List of cited references
1. Bechtold, U., Rabbani, N., Mullineaux, P. M., and Thornalley, P. J. Quantitative measurement of specific biomarkers for protein oxidation, nitration and glycation in Arabidopsis leaves // Plant J., 2009. V. 59. P. 661-671.
2. Bilova T, Lukasheva E, Brauch D, Greifenhagen U, Paudel G, Tarakhovskaya E, Frolova N, Mittasch J, Balcke GU, Tissier A, Osmolovskaya N, Vogt T, Wessjohann LA, Birkemeyer C, Milkowski C, Frolov A. A Snapshot of the Plant Glycated Proteome: Structural, Functional and Mechanistic Aspects // J. Biol. Chem., 2016. V. 291(14). P. 7621-7636.
3. Paudel G., Bilova T., Schmidt R., Greifenhagen U., Berger R., Tarakhovskaya E., Stöckhardt S., Balcke G.U., Humbeck K., Brandt W., Sinz A., Vogt T., Birkemeyer C., Wessjohann L., Frolov A. Osmotic stress is accompanied by protein glycation in Arabidopsis thaliana // J. Exp. Bot., 2016. V. 67 (22). P. 6283-6295.

Conference

ConferenceII Международный симпозиум "Молекулярные аспекты редокс-метаболизма растений" и Международной научной школы " Роль активных форм кислорода в жизни растений"
CountryRussian Federation
CityУфа
Period26/06/171/07/17
Internet address

Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology (miscellaneous)

Cite this

Bilova, T., Lukasheva, E., Paudel, G., Brauch, D., Tarakhovskaya, E., Kim, A., Vogt, T., Balcke, G. U., Birkemeyer, C., Wessjohann, L. A., & Frolov, A. (2017). ENVIRONMENTAL STRESS PROMOTES GLYCATION OF PLANT PROTEINS.. 26-28. Poster session presented at II Международный симпозиум "Молекулярные аспекты редокс-метаболизма растений" и Международной научной школы " Роль активных форм кислорода в жизни растений", Уфа, .