Protein Oxidataion in lipid and glucose toxicity

The aim of our studies is to provide insights into the role of protein oxidation in hepatocytes response to drugs, xenobiotics and genotoxic compounds.
We have developed a 3D cell culture-based model utilising hepatocarcinoma cell line and multicellular spheroids propagated in rotary bioreactors. We asses cell function by measuring physiological parameters such as glucose consumption, lipid uptake, lipid accumulation, ATP levels, protein content, glycogen levels and proliferation. We use proteomics to evaluate changes in protein expression and modification patterns and lipidomic analysis to follow lipid metabolism and accumulation. We have developed specific redox-proteomics approaches that allow us to follow S-nitrosylation and S-sulfenylation and protein carbonylation in biological samples.

Publications

1. Havelund, J. F.; Wojdyla, K.; Davies, M. J.; Jensen, O. N.; Møller, I. M.; Rogowska-Wrzesinska, A. (2017): A biotin enrichment strategy identifies novel carbonylated amino acids in proteins from human plasma. J Proteomics 156, 40-51. http://dx.doi.org/10.1016/j.jprot.2016.12.019.
2. Wojdyla, K.; Wrzesinski, K.; Williamson, J.; Fey, S. J.; Rogowska-Wrzesinska, A. (2016): Acetaminophen-induced S-nitrosylation and S-sulfenylation signalling in 3D cultured hepatocarcinoma cell spheroids. Toxicol Res (Camb) 5, 3, 905-920. http://dx.doi.org/10.1039/c5tx00469a.
3. Wojdyla, K.; Williamson, J.; Roepstorff, P.; Rogowska-Wrzesinska, A. (2015): The SNO/SOH TMT strategy for combinatorial analysis of reversible cysteine oxidations. J Proteomics 113, 415-434. http://dx.doi.org/10.1016/j.jprot.2014.10.015.
4. Wojdyla, K.; Rogowska-Wrzesinska, A. (2015): Differential alkylation-based redox proteomics--Lessons learnt. Redox Biol 6, 240-252. http://dx.doi.org/10.1016/j.redox.2015.08.005.
5. Nedić, O.; Rogowska-Wrzesinska, A.; Rattan, S. I. S. (2015): Standardization and quality control in quantifying non-enzymatic oxidative protein modifications in relation to ageing and disease: Why is it important and why is it hard? Redox Biol 5, 91-100. http://dx.doi.org/10.1016/j.redox.2015.04.001.
6. Augustyniak, E.; Adam, A.; Wojdyla, K.; Rogowska-Wrzesinska, A.; Willetts, R.; Korkmaz, A.; Atalay, M.; Weber, D.; Grune, T.; Borsa, C.; Gradinaru, D.; Chand Bollineni, R.; Fedorova, M.; Griffiths, H. R. (2015): Validation of protein carbonyl measurement: a multi-centre study. Redox Biol 4, 149-157. http://dx.doi.org/10.1016/j.redox.2014.12.014.
7. Rogowska-Wrzesinska, A.; Wojdyla, K.; Williamson, J.; Roepstorff, P. (2014): Measuring site occupancy: a new perspective on cysteine oxidation. Free Radic Biol Med 75 Suppl 1, S46-47. http://dx.doi.org/10.1016/j.freeradbiomed.2014.10.808.
8. Rogowska-Wrzesinska, A.; Wojdyla, K.; Nedić, O.; Baron, C. P.; Griffiths, H. R. (2014): Analysis of protein carbonylation--pitfalls and promise in commonly used methods. Free Radic Res 48, 10, 1145-1162. http://dx.doi.org/10.3109/10715762.2014.944868.
9. Møller, I. M.; Rogowska-Wrzesinska, A.; Rao, R. S. (2011): Protein carbonylation and metal-catalyzed protein oxidation in a cellular perspective. J Proteomics 74, 11, 2228-2242. http://dx.doi.org/10.1016/j.jprot.2011.05.004.